Assessment of gross and net mineralization rates of soil organic phosphorus – A review

The quantification of net soil organic P mineralization rates is hampered by the potentially rapid sorption of released phosphate. Here, isotopic dilution approaches to assess gross and net organic P mineralization rates under steady-state conditions are reviewed, including different analytical and numerical solutions to assess P transformation rates based on incubation experiments with ³²P- or ³³P-labeled soils. Non-isotopic approaches are also commented on. Published isotopic dilution studies show that isotopically exchangeable P during incubation can partly or even predominantly (20–90%) result from biological and biochemical rather than physicochemical processes. The relative contribution of biological and biochemical processes tends to be lower in arable soils than under grassland and forests and is negatively related to the availability of inorganic P and positively to concentrations of soil organic carbon. Typical basal gross organic P mineralization rates range between 0.1 and 2.5 mg P kg⁻¹ d⁻¹, but rates up to 12.6 mg P kg⁻¹ d⁻¹ have been observed in grassland and forest soils. The further partitioning of gross organic P mineralization remains uncertain, but a dominance of microbial immobilization and remineralization is likely under most conditions, at least during the initial weeks of incubation. Over longer time periods, the relative importance of mineralization of non-living soil organic P increases, with the contribution of extracellular hydrolysis remaining to be elucidated. This requires other approaches than enzyme activity assays, since measurements of phosphomonoesterase activity in soil render organic P mineralization rates that are one to two orders of magnitude greater than those determined by isotopic dilution. The numerical modeling approach will enable assessment of soil P transformation rates under non-steady-state conditions, where P fluxes are likely to be greater than under steady-state conditions. Ultimately, an improved understanding of the biological and biochemical processes in soil P dynamics may help to improve P management in agroecosystems.     

Immobilization and remineralization of N following addition of wheat straw into soil: determination of gross N transformation rates by N-ammonium isotope dilution technique

N dynamics in soil where wheat straw was incorporated were investigated by a soil incubation experiment using ¹⁵N-labelled nitrate or ¹⁵N-labelled wheat straw. The incubated soils were sampled after 7, 28, 54 days from the incorporation of wheat straw, respectively, and gross rates of N transformations including N remineralization and temporal changes in the amount of microbial biomass were determined.Following the addition of wheat straw into soils, rapid decrease of nitrate content in soil and increase of microbial biomass C and N occurred within the first week from onset of the experiment. Both the gross rates of mineralization and immobilization determined by ¹⁵N-ammonium isotope dilution technique were remarkably enhanced by the addition of wheat straw, and gradually decreased with time. Remineralization rate of N derived from ¹⁵N-labelled nitrate, and mineralization rate of N derived from ¹⁵N-labelled wheat straw was estimated by ¹⁵N isotope dilution technique using non-labelled ammonium. Remineralization rates of N derived from ¹⁵N-labelled nitrate were calculated to be 0.71 mg N kg⁻¹ d⁻¹ after 7 days, 0.55 mg N kg⁻¹ d⁻¹ after 28 days, and 0.29 mg N kg⁻¹ d⁻¹ after 54 days.Nearly 10% of the ¹⁵N-labelled N originally contained in the wheat straw was held in the microbial biomass irrespective of the sampling time. The amount of inorganic N in soil which was derived from ¹⁵N-labelled wheat straw ranged between 1.93 and 2.37 mg N kg⁻¹.Rates of N transformations in soil with ¹⁵N-labelled wheat straw were obtained by assuming that the k value was equal to the ¹⁵N abundance of biomass N, and the obtained values were considered to be valid.     

Basal organic phosphorus mineralization in soils under different farming systems

Soil organic P (P_o) mineralization plays an important role in soil P cycling. Quantitative information on the release of available inorganic P (P_i) by this process is difficult to obtain because any mineralized P_i gets rapidly sorbed. We applied a new approach to quantify basal soil P_o mineralization, based on ³³PO₄ isotopic dilution during 10 days of incubation, in soils differing in microbiological activity. The soils originated from a 20 years old field experiment, including a conventional system receiving exclusively mineral fertilizers (MIN), a bio-organic (ORG) and bio-dynamic (DYN) system. Indicators of soil microbiological activity, such as size and activity of the soil microbial biomass and phosphatase activity, were highest in DYN and lowest in MIN. In order to assess P_o hydrolysis driven by phosphatase in sterile soils, a set of soil samples was γ-irradiated. Basal P_o mineralization rates in non-irradiated samples were between 1.4 and 2.5 mg P kg⁻¹ day⁻¹ and decreased in the order DYN>ORG≥MIN. This is an amount lower, approximately equivalent to, or higher than water soluble P_i of MIN, ORG and DYN soils, respectively, but in every soil was less than 10% of the amount of P isotopically exchangeable during one day. This shows that physico-chemical processes are more important than basal mineralization in releasing plant available P_i. Organic P mineralization rates were higher, and differences between soils were more pronounced in γ-irradiated than in non-irradiated soils, with mineralization rates ranging from 2.2 to 4.6 mg P kg⁻¹ day⁻¹. These rates of hydrolysis, however, cannot be compared to those in non-sterile soils as they are affected by the release of cellular compounds, e.g. easily mineralizable P_o, derived from microbial cells killed by γ-irradiation.     

Gross sulphur mineralisation-immobilisation turnover in soil amended with plant residues

The rates of sulphur (S) released to and removed from the soil inorganic pool were estimated using the isotopic dilution technique. In an initial study fresh soil was mixed with combinations of two inorganic S levels (0 and 10 μg S g⁻¹ soil) and three plant residues (wheat straw, perennial ryegrass and oilseed rape) and followed over 32 days of incubation. As ³⁵S recovery was inadequate prior to day 2 and re-mineralisation of immobilised ³⁵S occurred after day 8 thereby invalidating the method, estimates of gross S transformation rates should be based on data sampled between days 2 and 8. In the main experiment 16 plant residues with ranges in S contents of 0.08-0.81%, C/S ratios of 50-604 and lignin content of 0.9-10.8 were mixed with soil and carrier-free ³⁵S label. Net turnover rates varied from 58% of S in Persian clover being immobilised to 76% of S in winter cress being mineralised within 5 days of incubation. Gross S mineralisation varied from 0.9-14.9 μg S g⁻¹ soil d⁻¹, whereas gross immobilisation only varied from 0.5 to 3.1 μg S g⁻¹ d⁻¹. Gross S immobilisation was strongly correlated to the C/S ratio of the plant material (P<0.001), whereas gross S mineralisation showed a weaker, but still significant, correlation with lignin content (P<0.05). The results indicate that immobilisation may predominantly have been a biological process in response to carbon addition while early mineralisation may have been dominated by the biochemical hydrolysis of organic sulphates in the residues. If attention is paid to the various constraints and limitations, isotopic pool dilution using ³⁵S offers a tool that may prove valuable in understanding and modelling soil S turnover.     

Gross nitrogen transformations in adjacent native and plantation forests of subtropical Australia

The impact of land-use change on soil nitrogen (N) transformations was investigated in adjacent native forest (NF), 53 y-old first rotation (1R) and 5 y-old second rotation (2R) hoop pine (Araucaia cunninghamii) plantations. The ¹⁵N isotope dilution method was used to quantify gross rates of N transformations in aerobic and anaerobic laboratory incubations. Results showed that the land-use change had a significant impact on the soil N transformations. Gross ammonification rates in the aerobic incubation ranged between 0.62 and 1.78 mg N kg⁻¹ d⁻¹, while gross nitrification rates ranged between 2.1 and 6.6 mg N kg⁻¹ d⁻¹. Gross ammonification rates were significantly lower in the NF and the 1R soils than in the 2R soils, however gross nitrification rates were significantly higher in the NF soils than in the plantation soils. The greater rates of gross nitrification found in the NF soil compared to the plantation soils, were related to lower soil C:N ratios (i.e. more labile soil N under NF). Nitrification was found to be the dominant soil N transformation process in the contrasting forest ecosystems. This might be attributed to certain site conditions which may favour the nitrifying community, such as the dry climate and tree species. There was some evidence to suggest that heterotrophic nitrifiers may undertake a significant portion of nitrification.     

Cyanobacterial inoculation of heated soils: effect on microorganisms of C and N cycles and on chemical composition in soil surface

Physiological groups of soil microorganisms, total C and N and available nutrients were investigated in four heated (350 °C, 1 h) soils (one Ortic Podsol over sandstone and three Humic Cambisol over granite, schist or limestone) inoculated (1.5 μg chlorophyll a g⁻¹ soil or 3.0 μg chlorophyll a g⁻¹ soil) with four cyanobacterial strains of the genus Oscillatoria, Nostoc or Scytonema and a mixture of them.Cyanobacterial inoculation promoted the formation of microbiotic crusts which contained a relatively high number of NH₄⁺-producers (7.4×10⁹ g⁻¹ crust), starch-mineralizing microbes (1.7×10⁸ g⁻¹ crust), cellulose-mineralizing microbes (1.4×10⁶ g⁻¹ crust) and NO₂⁻ and NO₃⁻ producers (6.9×10⁴ and 7.3×10³ g⁻¹ crust, respectively). These crusts showed a wide range of C and N contents with an average of 293 g C kg⁻¹ crust and 50 g N kg⁻¹ crust, respectively. In general, Ca was the most abundant available nutrient (804 mg kg⁻¹ crust), followed by Mg (269 mg kg⁻¹ crust), K (173 mg kg⁻¹ crust), Na (164 mg kg⁻¹ crust) and P (129 mg kg⁻¹ crust). There were close positive correlations among all the biotic and abiotic components of the crusts.Biofertilization with cyanobacteria induced great microbial proliferation as well as high increases in organic matter and nutrients in the surface of the heated soils. In general, cellulolytics were increased by four logarithmic units, amylolytics and ammonifiers by three logarithmic units and nitrifiers by more than two logarithmic units. C and N contents rose an average of 275 g C kg⁻¹ soil and 50 g N kg⁻¹ soil while the C:N ratio decreased up to 7 units. Among the available nutrients the highest increase was for Ca (315 mg kg⁻¹ soil) followed by Mg (189 mg kg⁻¹ soil), K (111 mg kg⁻¹ soil), Na (109 mg kg⁻¹ soil) and P (89 mg kg⁻¹ soil). Fluctuations of the microbial groups as well as those of organic matter and nutrients were positively correlated.The efficacy of inoculation depended on both the type of soil and the class of inoculum. The best treatment was the mixture of the four strains and, whatever the inoculum used, the soil over lime showed the most developed crust followed by the soils over schist, granite and sandstone. In the medium term there were not significant differences between the two inocula amounts tested.These results showed that inoculation of burned soils with alien N₂-fixing cyanobacteria may be a biotechnological means of promoting microbiotic crust formation, enhancing C and N cycling microorganisms and increasing organic matter and nutrient contents in heated soils.     

Rewetting and litter addition influence mineralisation and microbial communities in soils from a semi-arid intermittent stream

Nitrogen (N) and carbon (C) mineralisation are triggered by pulses of water availability in arid and semi-arid systems. Intermittent streams and their associated riparian communities are obvious ‘hot spots’ for biogeochemical processes in arid landscapes where water and often C are limiting. Stream landscapes are characterized by highly heterogeneous soils that may respond variably to rewetting. We used a laboratory incubation to quantify how N and C mineralisation in rewetted soils and sediments from an intermittent stream in the semi-arid Pilbara region of north-west Australia varied with saturation level and substrate addition (as ground Eucalyptus litter). Full (100%) saturation was defined as the maximum gravimetric moisture content (%) achieved in free-draining soils and sediments after rewetting, with 50% saturation defined as half this value. We estimated rates and amounts of N mineralised from changes in inorganic N and microbial respiration as CO₂ efflux throughout the incubation. In soils and sediments subject to 50% saturation, >90% of N mineralised accumulated within the first 7 d of incubation, compared to only 48% when soils were fully saturated (100% saturation). Mineralisation rates and microbial respiration were similar in riparian and floodplain soils, and channel sediments. N mineralisation rates in litter-amended soils and sediments (0.73 mg N kg⁻¹ d⁻¹) were only one-third that of unamended samples (3.04 mg N kg⁻¹ d⁻¹), while cumulative microbial respiration was doubled in litter-amended soils, suggesting N was more rapidly immobilized. Landscape position was less important in controlling microbial activity than soil saturation when water-filled pore space (% WFPS) was greater than 40%. Our results suggest that large pulses of water availability resulting in full soil saturation cause a slower release of mineralisation products, compared to small pulse events that stimulate a rapid cycle of C and N mineralisation-immobilization.     

Surface runoff phosphorus (P) loss in relation to phosphatase activity and soil P fractions in Florida sandy soils under citrus production

Phosphorus losses by surface runoff from agricultural lands have been of public concern due to increasing P contamination to surface waters. Five representative commercial citrus groves (C1-C5) located in South Florida were studied to evaluate the relationships between P fractions in soils, surface runoff P, and soil phosphatase activity. A modified Hedley P sequential fractionation procedure was employed to fractionate soil P. Soil P consisted of mainly organically- and Ca/Mg-bound P fractions. The organically-bound P (biological P, sum of organic P in the water, NaHCO₃ and NaOH extracts) was dominant in the acidic sandy soils from the C2 and C3 sites (18% and 24% of total soil P), whereas the Ca/Mg-bound P (HCl-extractable P) accounted for 45-60% of soil total P in the neutral and alkaline soils (C1, C4 and C5 soils). Plant-available P (sum of water and NaHCO₃ extractable P fractions) ranged from 27 to 61 mg P kg⁻¹ and decreased in the order of C3>C4>C1>C2>C5. The mean total P concentrations (TP) in surface runoff water samples ranged from 0.51 to 2.64 mg L⁻¹. Total P, total dissolved P (TDP), and PO₄³⁻-P in surface runoff were significantly correlated with soil biological P and plant-available P forms (p<0.01), suggesting that surface runoff P was directly derived from soil available P pools, including H₂O- and NaHCO₃- extractable inorganic P, water-soluble organic P, and NaHCO₃- and NaOH-extractable organic P fractions, which are readily mineralized by soil microorganisms and/or enzyme mediated processes. Soil neutral (55-190 mg phenol kg⁻¹ 3 h⁻¹) and natural (measured at soil pH) phosphatase activities (77-295 mg phenol kg⁻¹ 3 h⁻¹) were related to TP, TDP, and PO₄³⁻-P in surface runoff, and plant-available P and biological P forms in soils. These results indicate that there is a potential relationship between soil P availability and phosphatase activities, relating to P loss by surface runoff. Therefore, the neutral and natural phosphatase activities, especially the natural phosphatase activity, may serve as an index of surface runoff P loss potential and soil P availability.     

Priming effect of biuret addition on native soil N mineralisation under laboratory conditions

This study was carried out to quantify the priming effect of biuret on native soil nitrogen (N) mineralisation during a 112-day incubation. Addition of biuret (100 mg ¹⁵N-labelled biuret kg⁻¹ soil) increased the turnover rate constant of soil organic matter and had a positive priming effect on native soil N mineralisation in two soils. The additional mineralisation was 0.65% of the total soil N (equivalent to 47.1 kg N ha⁻¹) in a sandy loam soil and 0.62% of the soil N (equivalent to 46.5 kg N ha⁻¹) in a silt loam soil.     

Enzyme activities and microbial biomass in coastal soils of India

Soil salinity is a serious problem for agriculture in coastal regions, wherein salinity is temporal in nature. We studied the effect of salinity, in summer, monsoon and winter seasons, on microbial biomass carbon (MBC) and enzyme activities (EAs) of the salt-affected soils of the coastal region of the Bay of Bengal, Sundarbans, India. The average pH of soils collected from different sites, during different seasons varied from 4.8 to 7.8. The average organic C (OC) and total N (TN) content of the soils ranged between 5.2-14.1 and 0.6-1.4 g kg⁻¹, respectively. The electrical conductivity of the saturation extract (ECe) of soils, averaged over season, varied from 2.2 to 16.3 dSm⁻¹. The ECe of the soils increased five fold during the summer season (13.8 dSm⁻¹) than the monsoon season (2.7 dSm⁻¹). The major cation and anion detected were Na⁺ and Cl⁻, respectively. Seasonality exerted considerable effects on MBC and soil EAs, with the lowest values recorded during the summer season. The activities of β-glucosidase, urease, acid phosphatase and alkaline phosphatase were similar during the winter and monsoon season. The dehydrogenase activity of soils was higher in monsoon than in winter. Average MBC, dehydrogenase, β-glucosidase, urease, acid phosphatase and alkaline phosphatase activities of the saline soils ranged from 125 to 346 mg kg⁻¹ oven dry soil, 6-9.9 mg triphenyl formazan (TPF) kg⁻¹ oven dry soil h⁻¹, 18-53 mg p-nitro phenol (PNP) kg⁻¹ oven dry soil h⁻¹, 38-86 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 213-584 mg PNP kg⁻¹ oven dry soil h⁻¹ and 176-362 mg PNP g⁻¹ oven dry soil h⁻¹, respectively. The same for the non-saline soils were 274-446 mg kg⁻¹ oven dry soil, 8.8-14.4 mg TPF kg⁻¹ oven dry soil h⁻¹, 41-80 mg PNP kg⁻¹ oven dry soil h⁻¹, 89-134 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 219-287 mg PNP kg⁻¹ oven dry soil h⁻¹ and 407-417 mg PNP kg⁻¹ oven dry soil h⁻¹, respectively. About 48%, 82%, 48%, 63%, 40% and 48% variation in MBC, dehydrogenase activity, β-glucosidase activity, urease activity, acid phosphatase activity and alkaline phosphatase activity, respectively, could be explained by the variation in ECe of saline soils. Suppression of EAs of the coastal soils during summer due to salinity rise is of immense agronomic significance and needs suitable interventions for sustainable crop production.     

Seasonality of organic phosphorus mineralization in the rhizosphere of the native savanna grass, Trachypogon plumosus

Savanna ecosystems have low primary productivity, strong seasonality, and acid soils with low phosphorus (P) content. Organic P (Po) comprises around 50% of the total soil P and is plant-available only after mineralization. Rhizosphere processes mediated by plants, microorganisms and arbuscular mycorrhiza (AM) are important for plant P nutrition. We studied P transformation rates, Po-fractionation, acid phosphomonoesterase activity (APA), AM status, dehydrogenase activity (DHA), and bacterial and fungal plate counts in the rhizosphere of the native dominant grass Trachypogon plumosus. We collected samples from three acid savanna soils differing in order and P content (Entisol, Vertisol and Ultisol) at Estación Experimental La Iguana (Northeastern Venezuela) during the dry, rainy and transitional seasons over a 2-year-period. Less available Po fractions (moderately labile, moderately and highly resistant) seem to be involved in short-term P-cycling transformations as they significantly varied with season. During the rainy season plant P content (576-1160 mg P kg⁻¹ dry weight) and APA (44-200 mg PNP kg⁻¹ dry soil) were higher, while microbial number and activity (DHA) were lower. The higher P availability in the Entisol (6-9 mg P kg⁻¹ dry soil) resulted in a better plant nutritional status and inhibited APA. T. plumosus seems to be highly dependent on AM symbiosis (45-71% AM colonized root length, 0.6-8 AM spores g⁻¹ dry soil), especially during the rainy season. Po mineralization processes, mediated by biological associations in the rhizosphere, are crucial for understanding seasonal P-cycling and fertility in acid savanna soils.     

Rapid microbial phosphorus immobilization dominates gross phosphorus fluxes in a grassland soil with low inorganic phosphorus availability

Gross phosphorus (P) fluxes measured in isotopic dilution studies with ³³P labeled soils include the biological processes of microbial P immobilization, remineralization of immobilized P and mineralization of non-microbial soil organic P. In this approach, isotopic dilution due to physicochemical processes is taken into account. Our objectives were to assess the effect of inorganic P availability on gross P mineralization and immobilization in soil under permanent grassland, and to relate these fluxes to soil respiration, phosphatase activity and substrate availability as assessed by an enzyme addition method. We used soils from an 18-year-old grassland fertilization experiment near Zurich, Switzerland, that were collected in two treatments which differed only in the amount of mineral P applied (0 and 17 kg P ha⁻¹ yr⁻¹ in NK and NPK, respectively). Water-extractable phosphate was low (0.1 and 0.4 mg P kg⁻¹ soil in NK and NPK, while hexanol-labile (microbial) P was high (36 and 54 mg P kg⁻¹ soil in NK and NPK). Extremely fast microbial P uptake under P-limited conditions in NK necessitated the use of a microbial inhibitor when determining isotopic dilution due to physicochemical processes. At the higher inorganic P availability in NPK, however, isotopic exchange parameters were similar in the presence and absence of a microbial inhibitor. Phosphatase activity was higher in NK than in NPK, while soil respiration, water-extractable organic P and its enzyme-labile fraction were not affected by P status. Together, the results showed that inorganic P availability primarily affected microbial P immobilization which was the main component of gross P fluxes in both treatments. Gross P mineralization rates (8.2 and 3.1 mg P kg⁻¹ d⁻¹ for NK and NPK) during the first week were higher than reported in other studies on arable and forest soils and at least equal to isotopically exchangeable P due to physicochemical processes, confirming the importance of microbial processes in grassland soils.     

Phosphorus fractionation in lowland tropical rainforest soils in central Panama

Phosphorus availability is commonly assumed to limit productivity in lowland tropical rainforests, yet there is relatively little information on the chemical forms of soil phosphorus in such ecosystems. We used the Hedley sequential fractionation scheme to assess phosphorus chemistry in five soils supporting tropical rainforest on Barro Colorado Island, Republic of Panama. The soils represented a range of orders (Inceptisols, Alfisols, and Oxisols) formed on contrasting geological substrates and topography, but under uniform climate and vegetation. Total phosphorus in surface horizons ranged between 315 and 1114 mg P kg^− 1, being lowest on a soil derived from marine sediments and highest on soils derived from andesite. The majority of the phosphorus occurred in recalcitrant forms, although between 14% and 39% occurred as organic phosphorus. Readily-available phosphate, as extracted by anion-exchange membranes, occurred in small concentrations (4–13 mg P kg^− 1), although labile phosphorus, defined as phosphate extracted by anion-exchange membrane plus inorganic and organic phosphorus extracted by 0.5 M NaHCO₃, accounted for between 4.7% and 11.4% of the total soil phosphorus. Our results indicate a strong control of geology and topography on soil phosphorus in tropical rainforests, which may have important implications for understanding the diversity and distribution of plant species in such ecosystems. Further, some of the most common soils on Barro Colorado Island, including those on the 50 ha forest dynamics plot, are rich in phosphorus despite their relatively advanced stage of pedogenesis.     

Effects of decreasing water potential on gross ammonification and nitrification in an acid coniferous forest soil

Changes in the soil water regime, predicted as a consequence of global climate change, might influence the N cycle in temperate forest soils. We investigated the effect of decreasing soil water potentials on gross ammonification and nitrification in different soil horizons of a Norway spruce forest and tested the hypotheses that i) gross rates are more sensitive to desiccation in the Oa and EA horizon as compared to the uppermost Oi/Oe horizon and ii) that gross nitrification is more sensitive than gross ammonification. Soil samples were adjusted by air drying to water potentials from about field capacity to around −1.0 MPa, a range that is often observed under field conditions at our site. Gross rates were measured using the ¹⁵N pool dilution technique. To ensure that the addition of solute label to dry soils and the local rewetting does not affect the results by re-mineralization or preferential consumption of ¹⁵N, we compared different extraction and incubation times.T₀ times ranging from 10 to 300 min and incubation times of 48 h and 72 h did not influence the rates of gross ammonification and nitrification. Even small changes of water potential decreased gross ammonification and nitrification in the O horizon. In the EA horizon, gross nitrification was below detection limit and the response of the generally low rates of gross ammonification to decreasing water potentials was minor. In the Oi/Oe horizon gross ammonification and nitrification decreased from 37.5 to 18.3 mg N kg⁻¹ soil d⁻¹ and from 15.4 to 5.6 mg N kg⁻¹ soil d⁻¹ when the water potential decreased from field capacity to −0.8 MPa. In the Oa horizon gross ammonification decreased from 7.4 to 4.0 mg N kg⁻¹ soil d⁻¹ when the water potential reached −0.6 MPa. At such water potential nitrification almost ceased, while in the Oi/Oe horizon nitrification continued at a rather high level. Hence, only in the Oa horizon nitrification was more sensitive to desiccation than ammonification. Extended drought periods that might result from climate change will cause a reduction in gross N turnover rates in forest soils even at moderate levels of soil desiccation.     

Land use effects on gross nitrogen mineralization, nitrification, and N2O emissions in ephemeral wetlands

Stable ¹⁵N isotope dilution and tracer techniques were used in cultivated (C) and uncultivated (U) ephemeral wetlands in central Saskatchewan, Canada to: (1) quantify gross mineralization and nitrification rates and (2) estimate the relative proportion of N₂O emissions from these wetlands that could be attributed to denitrification versus nitrification-related processes. In-field incubation experiments were repeated in early May, mid-June and late July. Mean gross mineralization and nitrification rates (10.3 and 3.1 mg kg⁻¹ d⁻¹, respectively) did not differ between C and U wetlands on any given date. Despite these similarities, the mean NH₄⁺ pool size in the U wetlands (17.2 mg kg⁻¹) was two to three times that of the C wetlands (6.7 mg kg⁻¹) whereas the mean NO₃⁻ pool size in U wetlands (2.2 mg kg⁻¹) was less than half that of C wetlands (5.8 mg kg⁻¹). Mean N₂O emissions from the C wetlands decreased from 112.8 to 17.0 ng N₂O m² s⁻¹ from May to July, whereas mean U-wetland N₂O emissions ranged only from 31.8 to 51.1 ng N₂O m² s⁻¹ over the same period. This trend is correlated to water-filled pore space in C wetlands, demonstrating a soil moisture influence on emissions. Denitrification is generally considered the dominant emitter of N₂O under anaerobic conditions, but in the C wetlands, only 49% of the May emissions could be directly attributed to denitrification, decreasing to 29% in July. In contrast, more than 75% of the N₂O emissions from the U wetlands arose from denitrification of the soil NO₃⁻ pool throughout the season. These land use differences in emission sources and rates should be taken into consideration when planning management strategies for greenhouse gas mitigation.     

Assessing the toxic effect of nitrapyrin on nitrogen transformation in soil

A 28 d N transformation test was developed according to the OECD guideline 216. In the laboratory-based test, a suitable soil was amended with powdered plant meal as an organic N source. Soil samples of 1 kg treated with five concentrations of nitrapyrin (2-chloro-6-(trichloromethyl)-pyridine), in the range 1.0-100 mg kg⁻¹ dry weight were incubated for 28 d at 20±2 °C. A dose response was produced and the N mineralisation EC₅₀ (95% C.I.) for nitrapyrin was 3.1 (1.9-4.3) mg kg⁻¹ dry soil. The determined EC₅₀ was compared with literature figures for similar end points but using different methodology.     

Gross N transformation rates and net N mineralisation rates related to the C and N contents of soil organic matter fractions in grassland soils of different age

We investigated the relationship between soil organic matter (SOM) content and N dynamics in three grassland soils (0-10 and 10-20 cm depth) of different age (6, 14 and 50 y-old) with sandy loam textures. To study the distribution of the total C and N content the SOM was fractionated into light, intermediate and heavy density fractions of particulate macro-organic matter (150-2000 μm) and the 50-150 μm and <50 μm size fractions. The potential gross N transformation rates (mineralisation, nitrification, NH₄⁺ and NO₃⁻ immobilization) were determined by means of short-term, fully mirrored ¹⁵N isotope dilution experiments (7-d incubations). The long-term potential net N mineralisation and gross N immobilization rates were measured in 70-d incubations. The total C and N contents mainly tended to increase in the 0-10 cm layer with increasing age of the grassland soils. Significant differences in total SOM storage were detected for the long-term (50 y-old) conversion from arable land to permanent grassland. The largest relative increase in C and N contents had occurred in the heavy density fraction of the macro-organic matter, followed by the 50-150 and <50 μm fractions. Our results suggest that the heavy density fraction of the macro-organic matter could serve as a good indicator of early SOM accumulation, induced by converting arable land to permanent grassland. Gross N mineralisation, nitrification, and (long-term) gross N immobilization rates tended to increase with increasing age of the grasslands, and showed strong, positive correlations with the total C and N contents. The calculated gross N mineralisation rates (7-d incubations) and net N mineralisation rates (70-d incubations) corresponded with a gross N mineralisation of 643, 982 and 1876 kg N ha⁻¹ y⁻¹, and a net N mineralisation of 195, 208 and 274 kg N ha⁻¹ y⁻¹ in the upper 20 cm of the 6, 14 and 50 y-old grassland soils, respectively. Linear regression analysis showed that 93% of the variability of the gross N mineralisation rates could be explained by variation in the total N contents, whereas total N contents together with the C-to-N ratios of the <50 μm fraction explained 84% of the variability of the net N mineralisation rates. The relationship between long-term net N mineralisation rates and gross N mineralisation rates could be fitted by means of a logarithmic equation (net m=0.24Ln(gross m)+0.23, R²=0.69, P<0.05), which reflects that the ratio of gross N immobilization-to-gross N mineralisation tended to increase with increasing SOM contents. Microbial demand for N tended to increase with increasing SOM content in the grassland soils, indicating that potential N retention in soils through microbial N immobilization tends to be limited by C availability.     

Denitrification enzyme activity and potential of subsoils under grazed grasslands assayed by membrane inlet mass spectrometer

The importance of subsoil denitrification on the fate of agriculturally derived nitrate (NO₃) leached to groundwater is crucial for budgeting N in an ecosystem and for identifying areas where the risk of excess NO₃ is reduced. However, the high atmospheric background of di-nitrogen (N₂) causes difficulties in assessing denitrification enzyme activity (DEA) and denitrification potential (DP) in soils directly. Here, we apply Membrane Inlet Mass Spectrometry (MIMS) technique to investigate indirectly DEA and DP in soils by measuring N₂/Ar ratio changes in headspace water over soil. Soils were collected from 0-10, 15-25 and 60-70 cm depths of a grazed ryegrass and grass-clover. The samples were amended with helium-flushed deionized water containing ranges of NO₃ and carbon (glucose-C) and were incubated for six hours in the dark at 21 °C. The peaks for N₂/Ar ratio, declined with increasing soil depth, indicating a reduced substrate requirements to initiate DEA en-masse (15-30 mg NO₃-N alone or with 60-120 mg glucose-C, kg⁻¹ soil). The dissolved N₂O concentrations were very small (0.004-0.269 μg N kg⁻¹ soil) but responded well to the added N and C, showing a reduction in DEA with soil depth. In three separate studies, only subsoils were incubated for 3 days at 12 °C with 20-30 mg NO₃-N ± 40-60 mg glucose-C, kg⁻¹ soil. Denitrification capacity (DC, NO₃ only treatment) was not statistically different to the control (no amendment) within a land use (0.03-0.05 vs. 0.07-0.22 mg N kg⁻¹ soil d⁻¹), the highest being in ryegrass subsoils receiving groundwater. The DP was significantly (P < 0.0001) higher in subsoils under ryegrass than under grass-clover (0.50-0.71 vs. 1.15 mg N kg⁻¹ soil d⁻¹). The rates of DP (NO₃ + glucose-C) increased significantly (P < 0.0001) in unsaturated and saturated subsoils (0.92 and 2.19 mg N kg⁻¹ soil d⁻¹, respectively) of grass-clover, due to the higher reductive state resulting from the 10 day pre-incubation. Available C accelerated denitrification in soils and superseded the temporary elevation in oxidative state due to NO₃ addition. The substrates load differences between the land uses regulated the degree of denitrification rates. Results suggest that both dissolved N₂O measured by gas chromatography and N₂/Ar ratio measured by MIMS to indirectly determine DEA, and the latter to quantify total DC/DP in soils can be used. However, interference of oxygen in the MIMS system should be considered if available C is added or is naturally elevated in soil or groundwater.     

Microbial activity in pig slurry-amended soils under semiarid conditions

Soil amendment with manures from intensive animal industries is nowadays a common practice that may favorably or adversely affect several soil properties, including soil microbial activity. In this work, the effect of consecutive annual additions of pig slurry (PS) at rates of 30, 60, 90, 120 and 150 m³ ha⁻¹ y⁻¹ over a 4-year period on soil chemical properties and microbial activity was investigated and compared to that of an inorganic fertilization and a control (without amendment). Field plot experiment conducted under a continuous barley monoculture and semiarid conditions were used. Eight months after the fourth yearly PS and mineral fertilizer application (i.e. soon after the fourth barley harvest), surface soil samples (Ap horizon, 0-15 cm depth) from control and amended soils were collected and analysed for pH, electrical conductivity (EC), contents of total organic C, total N, available P and K, microbial biomass C, basal respiration and different enzymatic activities. The control soil had a slightly acidic pH (6.0), a small EC (0.07 dS m⁻¹), adequate levels of total N (1.2 g kg⁻¹) and available K (483 mg kg⁻¹) for barley growth, and small contents of total organic C (13.2 g kg⁻¹) and available P (52 mg kg⁻¹). With respect to the control and mineral fertilized soils, the PS-amended soils had greater pH values (around neutrality or slightly alkaline), electrical conductivities (still low) and contents of available P and K, and slightly larger total N contents. A significant decrease of total organic C was observed in soils amended at high slurry rate (12.3 g kg⁻¹). Compared with the control and mineral treatments, which produced almost similar results, the PS-amended soils were characterized by a higher microbial biomass C content (from 311 to 442 g kg⁻¹), microbial biomass C/total organic C ratio (from 2.3 to 3.6%) and dehydrogenase (from 35 to 173 μg INTF g⁻¹), catalase (from 5 to 24 μmol O₂ g⁻¹ min⁻¹), BAA-protease (from 0.7 to 1.9 μmol  g⁻¹ h⁻¹) and β-glucosidase (from 117 to 269 μmol PNP g⁻¹ h⁻¹) activities, similar basal respirations (from 48 to 77 μg C-CO₂ g⁻¹ d⁻¹) and urease activities (from 1.5 to 2.2 μmol  g⁻¹ h⁻¹), and smaller metabolic quotients (from 6.4 to 7.7 ng C-CO₂ μg⁻¹ biomass C h⁻¹) and phosphatese activities (from 374 to 159 μmol PNP g⁻¹ h⁻¹). For example, statistical analysis of experimental data showed that, with the exception of metabolic quotient and total organic C content, these effects generally increased with increasing cumulative amount of PS. In conclusion, cumulative PS application to soil over time under semiarid conditions may produce not only beneficial effects but also adverse effects on soil properties, such us the partial mineralization of soil organic C through extended microbial oxidation. Thus, PS should not be considered as a mature organic amendment and should be treated appropriately before it is applied to soil, so as to enhance its potential as a soil organic fertilizer.     

Heavy metal accumulation by two earthworm species and its relationship to total and DTPA-extractable metals in soils

The concentrations of Zn, Cd, Pb and Cu in earthworm tissues were compared with the total and DTPA-extractable contents of these heavy metals in contaminated soils. Samples were taken from a pasture polluted by waste from a metallurgic industry over 70 y ago. Three individuals of Aporrectodea caliginosa and Lumbricus rubellus and soil samples were collected at six points along a gradient of increasing pollution. Total metal contents of earthworms, soil, and metals extracted by DTPA from the soil were measured. Total heavy metal contents of the soils ranged from 165.7 to 1231.7 mg Zn kg⁻¹, 2.7 to 5.2 mg Cd kg⁻¹, 45.8 to 465.5 mg Pb kg⁻¹ and 30.0 to 107.5 mg Cu kg⁻¹. Their correlations with metals extracted by DTPA were highly significant. Contents of the metals in earthworm tissues were higher in A. caliginosa than in L. rubellus, with values ranging from 556 to 3381 mg Zn kg⁻¹, 11.6 to 102.9 mg Cd kg⁻¹, 1.9 to 182.8 mg Pb kg⁻¹ and 17.9 to 35.9 mg Cu kg⁻¹ in A. caliginosa, and from 667.9 to 2645 mg Zn kg⁻¹, 7.7 to 26.3 mg Cd kg⁻¹, 0.5 to 37.9 mg Pb kg⁻¹ and 16.0 to 37.6 mg Cu kg⁻¹ in L. rubellus, respectively. Correlations between body loads in earthworms with either total or DTPA-extractable contents of soil metals were significant, except for Cd in L. rubellus and Cu in A. caliginosa. Considering its simple analytical procedure, DTPA-extractable fraction may be preferable to total metal content as a predictor of bio-concentrations of heavy metals in earthworms. Biota-to-Soil Accumulation Factor (BSAF) of these four metals are Cd>Zn>Cu>Pb, with range of mean values between: Cd (6.18-17.02), Zn (1.95-7.91), Cu (0.27-0.89) and Pb (0.08-0.38) in A. caliginosa, and Cd (3.64-6.34), Zn (1.5-6.35), Cu (0.29-0.87) and Pb (0.04-0.13) in L. rubellus. The BSAF of Ca, Fe and Mn are Ca>Mn>Fe, with mean values of: Ca (0.46-1.31), Mn (0.041-0.111), Fe (0.017-0.07) in A. caliginosa and Ca (0.98-2.13), Mn (0.14-0.23), Fe (0.019-0.048) in L. rubellus, respectively. Results of principal component analysis showed that the two earthworm species differ in the pattern of metal bioaccumulation which is related to their ecological roles in contaminated soils.