Multi-site validation of a soil organic matter model for arable fields based on generally available input data

The paper describes a simplified carbon balance model, derived from the CANDY model, which works in annual time steps requiring only clay content, soil type of the German classification system “Reichsbodenschätzung”, average air temperature and rainfall as site characteristics, a value for organic carbon content as the initial value as well as crop yield and organic matter amendments as management data.The Candy Carbon Balance (CCB) model has been validated using a dataset from 40 long-term experiments situated in Central Europe including 391 treatments with a total number of 4794 C_org observations. Statistical measures to prove model validity were mean error (ME = − 0.001) and root mean square error (RMSE = 0.119). In addition a number of tests were performed to make sure that the model has no systematic error for different types of site conditions and management activities. After this successful validation the CCB model is considered applicable for advisory service for arable fields on a wide range of site conditions. Due to the poor representation of clay soils in this study some more model tests on these soils would be recommendable.     

Effects of potato–grain rotations on soil erosion, carbon dynamics and properties of rangeland sandy soils

The potential for wind erosion in South Central Colorado is greatest in the spring, especially after harvesting of crops such as potato (Solanum tuberosum L.) that leave small amounts of crop residue in the surface after harvest. Therefore it is important to implement best management practices that reduce potential wind erosion and that we understand how cropping systems are impacting soil erosion, carbon dynamics, and properties of rangeland sandy soils. We evaluate the effects of cropping systems on soil physical and chemical properties of rangeland sandy soils. The cropping system included a small grain–potato rotation. An uncultivated rangeland site and three fields that two decades ago were converted from rangeland into cultivated center-pivot-irrigation-sprinkler fields were also sampled. Plant and soil samples were collected in the rangeland area and the three adjacent cultivated sites. The soils at these sites were classified as a Gunbarrel loamy sand (Mixed, frigid Typic Psammaquent). We found that for the rangeland site, soil where brush species were growing exhibited C sequestration and increases in soil organic matter (SOM) while the bare soil areas of the rangeland are losing significant amounts of fine particles, nutrients and soil organic carbon (SOM-C) mainly due to wind erosion. When we compared the cultivated sites to the uncultivated rangeland, we found that the SOM-C and soil organic matter nitrogen (SOM-N) increased with increases in crop residue returned into the soils. Our results showed that even with potato crops, which are high intensity cultivated cropping systems, we can maintain the SOM-C with a rotation of two small grain crops (all residue incorporated) and one potato crop, or potentially increase the average SOM-C with a rotation of four small grain crops (all residue incorporated) and one potato crop. Erosion losses of fine silt and clay particles were reduced with the inclusion of small grains. Small grains have the potential to contribute to the conservation of SOM and/or sequester SOM-C and SOM-N for these rangeland systems that have very low C content and that are also losing C from their bare soils areas (40%). Cultivation of these rangelands using rotations with at least two small grain crops can reduce erosion and maintain SOM-C and increasing the number of small grain crops grown successfully in rotation above two will potentially contribute to C and N sequestration as SOM and to the sequestration of macro- and micro-nutrients.     

A comparison of the texture of grassland and eroded sandy soils from Shropshire, UK

In previous studies, periodic sampling of topsoils on runoff plots on sandy soils at the Hilton experimental site, Shropshire, UK, suggested erosion decreased the topsoil clay content and increased the coarse fraction. However, a comparison of soil and sediment properties suggested erosion selectively removed sand. Therefore, to cross-check the effects of erosion on soil properties, topsoil samples were collected from bare, eroded runoff plots and compared with samples from adjacent non-eroded grassland. Bare, eroded soil was stonier and particularly deficient in sand compared with grassed soil. Textural differences were very marked in the medium and coarse sands, especially the 0.5–1.0 mm fraction. On the basis of mean properties, the grassed soil was a very slightly stony loamy sand and the bare soil a slightly stony sandy loam. Soil organic matter was significantly less in the bare soils than the grassed soils and thus may have contributed to the higher erodibility of sands in bare soils.     

pH regulation of carbon and nitrogen dynamics in two agricultural soils

Soil pH is often hypothesized to be a major factor regulating organic matter turnover and inorganic nitrogen production in agricultural soils. The aim of this study was to critically test the relationship between soil pH and rates of C and N cycling, and dissolved organic nitrogen (DON), in two long-term field experiments in which pH had been manipulated (Rothamsted silty clay loam, pH 3.5-6.8; Woburn sandy loam, pH 3.4-6.3). While alteration of pH for 37 years significantly affected crop production, it had no significant effect on total soil C and N or indigenous mineral N levels. This implies that at steady state, increased organic matter inputs to the soil are balanced by increased outputs of CO₂. This is supported by the positive correlation between both plant productivity and intrinsic microbial respiration with soil pH. In addition, soil microbial biomass C and N, and nitrification were also significantly positively correlated with soil pH. Measurements of respiration following addition of urea and amino acids showed a significant decline in CO₂ evolution with increasing soil acidity, whilst glucose mineralization showed no response to pH. In conclusion, it appears that changes in soil pH significantly affect soil microbial activity and the rate of soil C and N cycling. The evidence suggests that this response is partially indirect, being primarily linked to pH induced changes in net primary production and the availability of substrates. In addition, enhanced soil acidity may also act directly on the functioning of the microbial community itself.     

Biochemical properties in managed grassland soils in a temperate humid zone: modifications of soil quality as a consequence of intensive grassland use

Although soil biochemical properties are considered to be good indicators of changes in soil quality, few studies have been made of the changes in biochemical properties brought about by anthropogenic disturbance of grassland ecosystems. In the present study, several biochemical properties were analysed in 31 grassland soils subjected to a high level of management, and the values obtained were compared with known values corresponding to native grasslands from the same region (Galicia, NW Spain). The 31 managed grasslands were divided into two groups (re-sown grasslands and improved grasslands) according to their management and past land use. The biochemical properties studied were: labile carbon, microbial biomass carbon, microbial respiration, metabolic quotient, net nitrogen mineralisation and the activities of dehydrogenase, catalase, phosphodiesterase, phosphomonoesterase, casein hydrolysing proteases, benzoyl arginamide (BAA)-hydrolysing proteases, urease, cellulase, ß-glucosidase, invertase and arylsulphatase. Managed grasslands exhibited lower values of soil biochemical properties than native grasslands. Three biochemical equilibrium equations were used to compare soil quality in managed and native grasslands. One of the equations did not show any significant difference between the groups of grassland soils considered. In contrast, two of the equations showed similar soil quality for improved and native grasslands, while re-sown grasslands exhibited a loss of soil quality when compared to native grassland soils.     

Mixing of different mineral soil layers by endogeic earthworms affects carbon and nitrogen mineralization

The effect of the endogeic earthworm species Octolasion tyrtaeum (Savigny) on decomposition of uniformly ¹⁴C-labelled lignin (lignocellulose) was studied in microcosms with upper mineral soil (Ah-horizon) from two forests on limestone, representing different stages of succession, a beech- and an ash-tree-dominated forest. Microcosms with and without lower mineral soil (Bw-horizon) were set-up; one O. tyrtaeum was added to half of them. It was hypothesised that endogeic earthworms stabilise lignin and the organic matter of the upper mineral soil by mixing with lower mineral soil of low C content. Cumulative C mineralization was increased by earthworms and by the addition of lower mineral soil. Effects of the lower mineral soil were more pronounced in the beech than in the ash forest. Cumulative mineralization of lignin was strongly increased by earthworms, but only in the beech soil (+24.6%). Earthworms predominantly colonized the upper mineral soil; mixing of the upper and lower mineral soils was low. The presence of lower mineral soil did not reduce the rates of decomposition of organic matter and lignin; however, the earthworm-mediated increase in mineralization was less pronounced in treatments with (+8.6%) than in those without (+14.1%) lower mineral soil. These results indicate that the mixing of organic matter with C-unsaturated lower mineral soil by endogeic earthworms reduced microbial decomposition of organic matter in earthworm casts.     

绰墩埋藏古水稻土中木质素特征研究

利用CuO氧化方法分析了苏州绰墩山埋藏古水稻土中木质素分解酚酸产物及特征,以此了解长期利用条件下水稻土中木质素的来源、保存及变化。通过比较相邻埋藏土壤剖面植稻和非植稻层次中木质素源的酚酸化合物特征,探讨水稻种植对土壤木质素结构特征的影响。结果表明,在埋藏古水稻土中木质素源的总酚酸含量在0.004～0.035 mg kg-1之间,显著低于现代水稻土(0.27～0.34 mg kg-1),而在埋藏土壤中,植稻与非植稻土壤层次中木质素无显著差异。埋藏古土壤中木质素源酚酸的组成(香草基类/紫丁香基类(S/V)及肉桂类/紫丁香基类(C/V)比分别为0.40～1.55、0.15～0.89),可证实此古土壤曾种植非木本被子植物;而S/V和C/V在植稻与非植稻土壤中没有显著差异,表明利用木质素特征难以区分同类(不同草本)有机质的具体来源。在埋藏古土壤中木质素碳占总有机碳比例很小,说明木质素在土壤中并非如通常所预期的容易被保存。     

Soil carbon dynamics in saline and sodic soils: a review

Soil salinity (high levels of water-soluble salt) and sodicity (high levels of exchangeable sodium), called collectively salt-affected soils, affect approximately 932 million ha of land globally. Saline and sodic landscapes are subjected to modified hydrologic processes which can impact upon soil chemistry, carbon and nutrient cycling, and organic matter decomposition. The soil organic carbon (SOC) pool is the largest terrestrial carbon pool, with the level of SOC an important measure of a soil's health. Because the SOC pool is dependent on inputs from vegetation, the effects of salinity and sodicity on plant health adversely impacts upon SOC stocks in salt-affected areas, generally leading to less SOC. Saline and sodic soils are subjected to a number of opposing processes which affect the soil microbial biomass and microbial activity, changing CO₂ fluxes and the nature and delivery of nutrients to vegetation. Sodic soils compound SOC loss by increasing dispersion of aggregates, which increases SOC mineralisation, and increasing bulk density which restricts access to substrate for mineralisation. Saline conditions can increase the decomposability of soil organic matter but also restrict access to substrates due to flocculation of aggregates as a result of high concentrations of soluble salts. Saline and sodic soils usually contain carbonates, which complicates the carbon (C) dynamics. This paper reviews soil processes that commonly occur in saline and sodic soils, and their effect on C stocks and fluxes to identify the key issues involved in the decomposition of soil organic matter and soil aggregation processes which need to be addressed to fully understand C dynamics in salt-affected soils.     

Incorporation of nitrogen from crop residues into light-fraction organic matter in soils with contrasting management histories

The proportion of N from crop residues entering the light-fraction organic matter (LFOM) pool was investigated in soils with contrasting soil organic matter and microbial characteristics arising from different management histories. A laboratory experiment was conducted in which ¹⁵N-labelled sugar beet, Brussels sprout or ryegrass shoots, which possessed a range of C/N contents, and hence different biochemical qualities, were incorporated into a sandy–loam soil collected from within a field (FC) or from the field margin (FM). Amounts of C and N incorporated into LFOM were determined after 112 days. The FC and FM soils had organic C contents of 0.9% and 2.5%, respectively. Addition of crop residues increased total LFOM N content and reduced its C/N in FC soil but had no effect on total LFOM N or its C/N in FM soil. Ryegrass incorporation into FC was the only treatment in which there was a net increase in LFOM C. Isotopic analysis indicated that more crop-residue-derived N became incorporated into the LFOM N pool in FM relative to FC soil, with per cent crop residue N incorporated ranging from 25.9% to 35.3% in FC and between 38.9 and 68.5 in FM. Incorporation of crop residues had a positive priming effect on pre-existing LFOM N in FM but not FC soil. We conclude that the characteristics of plant material, together with differences in soil organic matter and microbiology resulting from contrasting management, determined the amount of crop residue C and N incorporated into both HFOM and LFOM.     

The impact of a low humus level in arable soils on microbial properties, soil organic matter quality and crop yield

 In arable soils in Schleswig-Holstein (Northwest Germany) nearly 30% of the total organic C (TOC) stored in former times in the soil has been mineralized in the last 20 years. Microbial biomass, enzyme activities and the soil organic matter (SOM) composition were investigated in order to elucidate if a low TOC level affects microbial parameters, SOM quality and crop yield. Microbial biomass C (C_mic) and enzyme activities decreased in soils with a low TOC level compared to soils with a typical TOC level. The decrease in the C_mic/TOC ratio suggested low-level, steady-state microbial activity. The SOM quality changed with respect to an enrichment of initial litter compounds in the top soil layers with a low TOC level. Recent management of the soils had not maintained a desirable level of humic compounds. However, we found no significant decrease in crop yield. We suggest that microbial biomass and dehydrogenase and alkaline phosphatase activities are not necessarily indicators of soil fertility in soils with a high fertilization level without forage production and manure application. Received: 12 December 1997     

Depth distribution of soil organic C and N after long-term soybean cropping in Texas

Crop management practices have potential to enhance subsoil C and N sequestration in the southern U.S., but effects may vary with tillage regime and cropping sequence. The objective of this study was to determine the impacts of tillage and soybean cropping sequence on the depth distribution of soil organic C (SOC), dissolved organic C (DOC), and total N after 20 years of treatment imposition for a silty clay loam soil in central Texas. A continuous soybean monoculture, a wheat–soybean doublecrop, and a sorghum–wheat–soybean rotation were established under both conventional (CT) and no tillage (NT). Soil was sampled after soybean harvest and sectioned into 0–5, 5–15, 15–30, 30–55, 55–80, and 80–105 cm depth intervals. Both tillage and cropping intensity influenced C and N dynamics in surface and subsurface soils. No tillage increased SOC, DOC, and total N compared to CT to a 30 cm depth for continuous soybean, but to 55 cm depths for the more intensive sorghum–wheat–soybean rotation and wheat–soybean doublecrop. Averaged from 0 to 105 cm, NT increased SOC, DOC, and total N by 32, 22, and 34%, respectively, compared to CT. Intensive cropping increased SOC and total N at depths to 55 cm compared to continuous soybean, regardless of tillage regime. Continuous soybean had significantly lower SOC (5.3 g kg⁻¹) than sorghum–wheat–soybean (6.4 g kg⁻¹) and wheat–soybean (6.1 g kg⁻¹), and 19% lower total N than other cropping sequences. Dissolved organic C was also significantly higher for sorghum–wheat–soybean (139 mg C kg⁻¹) than wheat–soybean (92 mg C kg⁻¹) and continuous soybean (100 mg C kg⁻¹). The depth distribution of SOC, DOC, and total N indicated treatment effects below the maximum tillage depth (25 cm), suggesting that roots, or translocation of dissolved organic matter from surface soils, contributed to higher soil organic matter levels under NT than CT in subsurface soils. High-intensity cropping sequences, coupled with NT, resulted in the highest soil organic matter levels, demonstrating potential for C and N sequestration for subsurface soils in the southern U.S.     

Quantifying surface and subsurface cast production by earthworms under controlled laboratory conditions

Earthworm casts, formed when organic substrates and soil minerals pass through the digestive tract, may protect soil organic matter from biological degradation if they persist in the soil. Yet, the stability of casts is affected by their location in the soil profile because surface casts are exposed to more disruptive forces (wetting-drying, freezing-thawing) than subsurface casts. It is not known whether environmental conditions affect the proportions of surface and subsurface casts produced by earthworms. This study investigated how surface and subsurface cast production by juveniles of Aporrectodea spp. and Lumbricus spp. was affected by temperature. Two juveniles of Aporrectodea spp. or Lumbricus spp. were added to plexiglass chambers filled with soil, and five replicate chambers were incubated in the dark at 5°C, 10°C, 15°C or 20°C for 1 week. Most of the casts produced by Aporrectodea spp. and Lumbricus spp. were surface casts, with <10% of casts deposited below the soil surface. The earthworms studied produced more casts, and a greater proportion of surface casts, as the soil temperature increased. These results can be used to estimate the quantity of surface and subsurface casts produced by earthworm populations under field conditions and determine the susceptibility of cast-associated organic matter to decomposition in the medium- to long-term.     

Biochemical properties and biodegradation of dissolved organic matter from soils

Various biologically mediated processes are involved in the turnover of dissolved organic matter (DOM) in soil; however, relatively little is known about the dynamics of either the microbial community or the individual classes of organic molecules during the decomposition of DOM. We examined the net loss of DOC, the mineralisation of C to CO₂ and the degradation of DOC from six different soils by soil microorganisms. We also quantified the changes in the concentrations of protein, carbohydrate and amino acid C during microbial biodegradation. Over a 70-day incubation period at 20°C, the mineralisation of DOC to CO₂ was described by a double exponential model with a labile pool (half-life, 3–8 days) and a stable pool (half-life, 0.4–6 years). However, in nearly all cases, the mass loss of DOC exceeded the C released as CO₂ with significant deviations from the double exponential model. Comparison of mass DOC loss, CO₂ production and microbial cell counts, determined by epifluorescence microscopy, showed that a proportion of the lost DOC mass could be accounted for by microbial assimilation. Carbohydrate and protein C concentrations fluctuated throughout the incubation with a net change of between 3 to 13 and −30 to 22.4% initial DOC, respectively. No amino acid C was detected during the incubation period (level of detection, 0.01 mg C l⁻¹).     

The carbon we do not see—the impact of low molecular weight compounds on carbon dynamics and respiration in forest soils: a review

Dissolved organic matter (DOM), typically quantified as dissolved organic carbon (DOC), has been hypothesized to play many roles in pedogenesis and soil biogeochemical cycles, however, most research to date concerning forest soils has focussed on the high molecular weight (HMW) components of this DOM. This review aims to assess the role of low molecular weight (LMW) DOM compounds in the C dynamics of temperate and boreal forest soils focussing in particular on organic acids, amino acids and sugars. The current knowledge of concentrations, mineralization kinetics and production rates and sources in soil are summarised. We conclude that although these LMW compounds are typically maintained at very low concentrations in the soil solution (<50 μM), the flux through this pool is extremely rapid (mean residence time 1-10 h) due to continued microbial removal. Due to this rapid flux through the soil solution pool and mineralization to CO₂, we calculate that the turnover of these LMW compounds may contribute substantially to the total CO₂ efflux from the soil. Moreover, the production rates of these soluble transitory compounds could exceed HMW DOM production. The possible impact of climate change on the behaviour of LMW compounds in soil is also discussed.     

The soil carbon dilemma: Shall we hoard it or use it?

Rapidly rising concentrations of atmospheric CO₂ have prompted a flurry of studies on soils as potential carbon (C) ‘sinks’. Sequestering C in soils is often seen as a ‘win-win’ proposition; it not only removes excess CO₂ from the air, but also improves soils by augmenting organic matter, an energy and nutrient source for biota. But organic matter is most useful, biologically, when it decays. So we face a dilemma: can we both conserve organic matter and profit from its decay? Or must we choose one or the other? In this essay, I contemplate the merits, first of building soil C and then of decaying (losing) it, partly from a historical perspective. I then consider the apparent trade-off between accrual and decay, and reflect on how the dilemma might be resolved or assuaged. These fledgling thoughts, offered mostly to stir more fruitful debate, include: finding ways to increase C inputs to soil; seeking to optimize the timing of decay; and understanding better, from an ecosystem perspective, the flows of C, rather than only the stocks. Carbon sequestration is a sound and worthy goal. But soil organic matter is far more than a potential tank for impounding excess CO₂; it is a relentless flow of C atoms, through a myriad of streams—some fast, some slow—wending their way through the ecosystem, driving biotic processes along the way. Now, when we aim to regain some of the C lost, we may need new ways of thinking about soil C dynamics, and tuning them for the services expected of our ecosystems. This objective, perhaps demanding more biology along with other disciplines, is especially urgent when we contemplate the stresses soon to be imposed by coming global changes.     

Labile, recalcitrant, and inert organic matter in Mediterranean forest soils

The biochemical quality of soil organic matter (SOM) was studied in various profiles under Quercus rotundifolia Lam. stands on calcareous parent material. Special attention was paid to the question of how biochemical quality is affected by position within the soil profile (upper versus lower horizons). The following global SOM characteristics were investigated: (a) overall recalcitrance, using hydrolysis with either hydrochloric or sulphuric acid; (b) hydrolyzable carbohydrates and polyphenolics; (c) extractability by hot water and quality of the extract; and (d) abundance of inert forms of SOM: charcoal and soot-graphite. The recalcitrance of soil organic carbon (OC) decreases with depth, following the order: H horizons>A horizons>B horizons. In contrast, the recalcitrance of nitrogen is roughly maintained with depth. The ratio carbohydrate C to total OC increases from H to B horizons, due to the increasing importance of cellulosic polysaccharides in B horizons, whereas other carbohydrates are maintained throughout the soil profile at a relatively constant level, 12-15% of the total OC in the horizon. Whereas the quality of the hydrolyzable carbon (measured by the carbohydrate to polyphenolic C ratio) decreases with depth from H to B horizons, the quality of the hot-water extractable organic matter is much higher in B horizons than in A or H horizons. The relative importance of both charcoal and soot-graphitic C and N tends to increase with depth. The ratio black/total is usually higher for N than for C, a result that suggests that inert SOM may represent a relevant compartment in the nitrogen cycle. Overall, our data suggest that in Mediterranean forest soils the organic matter in B horizons could be less stable than often thought.     

Estimating soil labile organic carbon and potential turnover rates using a sequential fumigation-incubation procedure

Labile carbon is the fraction of soil organic carbon with most rapid turnover times and its oxidation drives the flux of CO₂ between soils and atmosphere. Available chemical and physical fractionation methods for estimating soil labile organic carbon are indirect and lack a clear biological definition. We have modified the well-established Jenkinson and Powlson's fumigation-incubation technique to estimate soil labile organic carbon using a sequential fumigation-incubation procedure. We define soil labile organic carbon as the fraction of soil organic carbon degradable during microbial growth, assuming that labile organic carbon oxidizes according to a simple negative exponential model. We used five mineral soils and a forest Oa horizon to represent a wide range of organic carbon levels. Soil labile organic carbon varied from 0.8 mg/g in an Entisol to 17.3 mg/g in the Oa materials. Potential turnover time ranged from 24 days in an Alfisol to 102 days in an Ultisol. Soil labile organic carbon contributed from 4.8% in the Alfisol to 11.1% in the Ultisol to the total organic carbon. This new procedure is a relatively easy and simple method for obtaining indices for both the pool sizes and potential turnover rates of soil labile organic carbon and provides a new approach to studying soil organic carbon.     

Carbon and nitrogen sequestration and soil aggregation under sorghum cropping sequences

Management practices, such as no tillage (NT) and intensive cropping, have potential to increase C and N sequestration in agricultural soils. The objectives of this study were to investigate the impacts of conventional tillage (CT), NT, and cropping intensity on soil organic C (SOC) and N (SON) sequestration and on distribution within aggregate-size fractions in a central Texas soil after 20 years of treatment imposition. Tillage regime and cropping sequence significantly impacted both SOC and SON sequestration. At 0–5 cm, NT increased SOC storage compared to CT by 33% and 97% and SON storage by 25% and 117% for a sorghum/wheat/soybean (SWS) rotation and a continuous sorghum monoculture, respectively. Total SOC and SON storage at both 0–5 and 5–15 cm was greater for SWS than continuous sorghum regardless of tillage regime. The majority of SOC and SON storage at 0–5 cm was observed in 250-m to 2-mm aggregates, and at 5–15 cm, in the >2-mm and 250-m to 2-mm fractions. Averaged across cropping sequences at 0–5 cm, NT increased SOC storage compared to CT by 212%, 96%, 0%, and 31%, and SON storage by 122%, 92%, 0%, and 37% in >2-mm, 250-m to 2-mm, 53- to 250-m, and <53-m aggregate-size fractions. No tillage and increased cropping intensity improved soil fertility by increasing soil organic matter levels and potential nutrient supply to crops.     

Urease activity in two cultivated and non-cultivated arid soils

Summary Since urease activity in soil is believed to be relatively constant, the present study was designed to examine the effects of incubation, soil depth and the effect of cultivation on the persistence of urease activity in arid soils. Two soils were used, a Harkey (coarse, silty, mixed, calcareous, thermic, Typic Torrifluvent) and a Saneli (Clayey over sandy skeletal, montmorillonitic, calcareous, Vertic Torrifluvent), each consisting of a cultivated field and a non-cultivated roadbed site. Urease activity was much lower and more varable in the roadbed soils (40 years without cultivation) than in the cultivated field soils. Pre-incubation for 24 h with urea (with toluene) and without urea (without toluene) greatly reduced the total urease activity in all cases in relation to cell free urease activity (with toluene). Urease activity in the two field soils decreased slightly with profile depth but the decrease was greatest below the plow depth (33 cm). Protease activity or some inactivation processes must have lowered the urease content since there was substantially reduced urease activity after most pre-incubations. The extent of the urease activity decrease was so great that the addition of urea would have been required to increase the production of urease enzyme.