Protection of soil carbon by microaggregates within earthworm casts

Earthworms are known to play a role in aggregate formation and soil organic matter (SOM) protection. However, it is still unclear at what scale and how quickly earthworms manage to protect SOM. We investigated the effects of Aporrectodea caliginosa on aggregation and aggregate-associated C pools using ¹³C-labeled sorghum (Sorghum bicolor (L.) Moench) leaf residue. Two incubations were set up. The first incubation consisted of soil samples crushed <250 μm to break up all macroaggregates with three treatments: (i) control soil; (ii) soil+¹³C-labeled residue and (iii) soil+¹³C-labeled residue+earthworms. Earthworms were added after 8 d and 12 d (days) later, aggregate size distribution was measured together with total C and ¹³C in each aggregate fraction. A second incubation was made to assay protected versus unprotected total C and ¹³C from 21-d laboratory incubations of intact and crushed large (>2000 μm) and small (250-2000 μm) macroaggregates and microaggregates (53-250 μm). Eight different pools of aggregate-associated C were quantified: (1) and (2) unprotected C pools in large and small macroaggregates, (3) unprotected C pools in microaggregates, (4) and (5) protected C pools in large and small macroaggregates, (6) protected C pool in microaggregates, and (7) and (8) protected C pools in microaggregates within large and small macroaggregates. In the presence of earthworms, a higher proportion of large macroaggregates was newly formed and these aggregates contained more C and ¹³C compared to bulk soil. There were no significant differences between the samples with or without earthworms in the C pool-sizes protected by macroaggregates, microaggregates or microaggregates within small macroaggregates. However, in the presence of earthworms, the C protected by microaggregates within large macroaggregates was a significant pool and 22% of this C pool was newly added C. In conclusion, these results clearly indicate the direct involvement of earthworms in providing protection of soil C in microaggregates within large macroaggregates leading to a possible long-term stabilization of soil C.     

Storage and dynamics of carbon and nitrogen in soil physical fractions following woody plant invasion of grassland

Woody plant invasion of grasslands is prevalent worldwide. In the Rio Grande Plains of Texas, subtropical thorn woodlands dominated by C₃ trees/shrubs have been replacing C₄ grasslands over the past 150 yr, resulting in increased soil organic carbon (SOC) storage and concomitant increases in soil total nitrogen (STN). To elucidate mechanisms of change in SOC and STN, we separated soil organic matter into specific size/density fractions and determined the concentration of C and N in these fractions. Soils were collected from remnant grasslands (Time 0) and woody plant stands (ages 10-130 yr). Rates of whole-soil C and N accrual in the upper 15 cm of the soil profile averaged 10-30 g C m⁻² yr⁻¹ and 1-3 g N m⁻² yr⁻¹, respectively, over the past 130 yr of woodland development. These rates of accumulation have increased soil C and N stocks in older wooded areas by 100-500% relative to remnant grasslands. Probable causes of these increased pool sizes include higher rates of organic matter production in wooded areas, greater inherent biochemical resistance of woody litter to decomposition, and protection of organic matter by stabilization within soil macro- and microaggregates. The mass proportions of the free light fraction (<1.0 g cm⁻³) and macroaggregate fraction (>250 μm) increased linearly with time following woody plant invasion of grassland. Conversely, the mass proportions of free microaggregate (53-250 μm) and free silt+clay (<53 μm) fractions decreased linearly with time after woody invasion, likely reflecting stabilization of these fractions within macroaggregate structures. Carbon and N concentrations increased in all soil fractions with time following woody invasion. Approximately half of the C and N accumulated in free particulate organic matter (POM) fractions, while the remainder accrued in stable macro- and microaggregate structures. Soil C/N ratios indicated that the organic C associated with POM and macroaggregates was of more recent origin (less decomposed) than C associated with the microaggregate and silt+clay fractions. Because grassland-to-woodland conversion has been geographically extensive in grassland ecosystems worldwide during the past century, changes in soil C and N storage and dynamics documented here could have significance for global cycles of those elements.     

Impacts of different N management regimes on nitrifier and denitrifier communities and N cycling in soil microenvironments

Real-time quantitative PCR assays, targeting part of the ammonia monooxygenase (amoA), nitrous oxide reductase (nosZ), and 16S rRNA genes were coupled with ¹⁵N pool dilution techniques to investigate the effects of long-term agricultural management practices on potential gross N mineralization and nitrification rates, as well as ammonia-oxidizing bacteria (AOB), denitrifier, and total bacterial community sizes within different soil microenvironments. Three soil microenvironments [coarse particulate organic matter (cPOM; >250 μm), microaggregate (53-250 μm), and silt-and-clay fraction (<53 μm)] were physically isolated from soil samples collected across the cropping season from conventional, low-input, and organic maize-tomato systems (Zea mays L.-Lycopersicum esculentum L.). We hypothesized that (i) the higher N inputs and soil N content of the organic system foster larger AOB and denitrifier communities than in the conventional and low-input systems, (ii) differences in potential gross N mineralization and nitrification rates across the systems correspond with AOB and denitrifier abundances, and (iii) amoA, nosZ, and 16S rRNA gene abundances are higher in the microaggregates than in the cPOM and silt-and-clay microenvironments. Despite 13 years of different soil management and greater soil C and N content in the organic compared to the conventional and low-input systems, total bacterial communities within the whole soil were similar in size across the three systems (∼5.15 × 10⁸ copies g⁻¹ soil). However, amoA gene densities were ∼2 times higher in the organic (1.75 × 10⁸ copies g⁻¹ soil) than the other systems at the start of the season and nosZ gene abundances were ∼2 times greater in the conventional (7.65 × 10⁷ copies g⁻¹ soil) than in the other systems by the end of the season. Because organic management did not consistently lead to larger AOB and denitrifier communities than the other two systems, our first hypothesis was not corroborated. Our second hypothesis was also not corroborated because canonical correspondence analyses revealed that AOB and denitrifier abundances were decoupled from potential gross N mineralization and nitrification rates and from inorganic N concentrations. Our third hypothesis was supported by the overall larger nitrifier, denitrifier, and total bacterial communities measured in the soil microaggregates compared to the cPOM and silt-and-clay. These results suggest that the microaggregates are microenvironments that preferentially stabilize C, and concomitantly promote the growth of nitrifier and denitrifier communities, thereby serving as potential hotspots for N₂O losses.     

Toxicological effects of TiO2 and ZnO nanoparticles in soil on earthworm Eisenia fetida

Nanoparticles (NPs) of TiO₂ and ZnO are receiving increasing attention due to their widespread applications. To evaluate their toxicities to the earthworm Eisenia fetida (Savigny, 1826) in soil, artificial soil systems containing distilled water, 0.1, 0.5, 1.0 or 5.0 g kg⁻¹ of NPs were prepared and earthworms were exposed for 7 days. Contents of Zn and Ti in earthworm, activities of antioxidant enzymes, DNA damage to earthworm, activity of cellulase and damage to mitochondria of gut cells were investigated after acute toxicity test. The results from response of the antioxidant system combined with DNA damage endpoint (comet assay) indicated that TiO₂ and ZnO NPs could induce significant damage to earthworms when doses were greater than 1.0 g kg⁻¹. We found that Ti and Zn, especially Zn, were bioaccumulated, and that mitochondria were damaged at the highest dose in soil (5.0 g kg⁻¹). The activity of cellulase was significantly inhibited when organisms were exposed to 5.0 g kg⁻¹ of ZnO NPs. Our study demonstrates that both TiO₂ and ZnO NPs exert harmful effects to E. fetida when their levels are higher than 1.0 g kg⁻¹ in soil and that toxicity of ZnO NPs was higher than TiO₂.     

Controls over pathways of carbon efflux from soils along climate and black spruce productivity gradients in interior Alaska

Small changes in C cycling in boreal forests can change the sign of their C balance, so it is important to gain an understanding of the factors controlling small exports like water-soluble organic carbon (WSOC) fluxes from the soils in these systems. To examine this, we estimated WSOC fluxes based on measured concentrations along four replicate gradients in upland black spruce (Picea mariana [Mill.] BSP) productivity and soil temperature in interior Alaska and compared them to concurrent rates of soil CO₂ efflux. Concentrations of WSOC in organic and mineral horizons ranged from 4.9 to 22.7 g C m⁻² and from 1.4 to 8.4 g C m⁻², respectively. Annual WSOC fluxes (4.5-12.0 g C m⁻² y⁻¹) increased with annual soil CO₂ effluxes (365-739 g C m⁻² y⁻¹) across all sites (R²=0.55, p=0.02), with higher fluxes occurring in warmer, more productive stands. Although annual WSOC flux was relatively small compared to total soil CO₂ efflux across all sites (<3%), its relative contribution was highest in warmer, more productive stands which harbored less soil organic carbon. The proportions of relatively bioavailable organic fractions (hydrophilic organic matter and low molecular weight acids) were highest in WSOC in colder, low-productivity stands whereas the more degraded products of microbial activity (fulvic acids) were highest in warmer, more productive stands. These data suggest that WSOC mineralization may be a mechanism for increased soil C loss if the climate warms and therefore should be accounted for in order to accurately determine the sensitivity of boreal soil organic C balance to climate change.     

Changes in soil organic carbon dynamics in an Eastern Chinese coastal wetland following invasion by a C4 plant Spartina alterniflora

The exotic C₄ grass Spartina alterniflora was intentionally introduced to tidal coastal wetlands in Jiangsu province of China in 1982. Since then it has rapidly replaced the native C₃ plant Suaeda salsa, becoming one of the dominant vegetation types in the coastal wetlands of China. Although plant invasion can change soil organic carbon (SOC) storage, little is known about how plant invasion influences C storage within soil fractions. We investigated how S. alterniflora invasion across an 8, 12 and 14-year chronosequence affected SOC and soil nitrogen (N), using soil fractionation and stable δ¹³C isotope analyses. SOC and N concentrations at 0-10 cm depth in S. alterniflora soil increased during the S. alterniflora invasion chronosequence, ranging from 3.67 to 4.90 g C kg⁻¹ soil, and from 0.307 to 0.391 g N kg⁻¹ soil. These were significantly higher than the values in the Suaeda salsa community, by 27.0-69.6% for SOC, and 21.8-55.2% for total N. The S. alterniflora-derived SOC varied from 0.40 to 0.92 g C kg⁻¹ according to mixing calculations, assuming the two possible SOC sources of S. alterniflora and S. salsa, and accounted for 10.8-18.7% of total SOC in the colonized soils. The estimated accumulative rate of SOC from C₄ (S. alterniflora) was 64.1 C kg⁻¹ soil year⁻¹ and from C₃ sources was 78.1 mg C kg⁻¹. The concentration of S. alterniflora-derived SOC significantly decreased from coarse fraction to fine fraction, and linearly increased as the period of S. alterniflora invasion increased. The highest accumulative rate of SOC from a C₄ source occurred in macroaggregates, while the highest rate from C₃ was in microaggregates. The storage of SOC derived from S. alterniflora in the macroaggregates was 0.27-0.44 g C kg⁻¹ soil, accounting for 43.1-49.1% of the total C₄derived SOC in the soil. Our results suggest that S. alterniflora invasion in coastal wetlands could facilitate SOC storage, because of the high potential for accumulation of the C which has been newly derived from S. alterniflora litter and roots.     

Rapid incorporation of carbon from fresh residues into newly formed stable microaggregates within earthworm casts

Earthworms play an important role in protecting carbon in the soil, but the exact influence of their activity on the distribution and protection of C is still poorly understood. We investigated the effect of earthworms on the formation of stable microaggregates inside newly formed macroaggregates and the distribution of C in them. We crushed (< 250 µm) soil, and subjected it to three treatments: (i) soil + ¹³C‐labelled residue + earthworms (these added after 8 days' incubation), (ii) soil + ¹³C‐labelled residue, and (iii) control (no additions), and then incubated it for 20 days. At the end, we measured the aggregate size distribution, total C and ¹³C, and we isolated microaggregates (53–250 µm) from macroaggregates (> 250 µm) formed. The ¹³C in fine particulate organic matter between and within the microaggregates was determined. Earthworms helped to form large macroaggregates (> 2000 µm). These large macroaggregates contained four times more stable microaggregates than those from samples without earthworms. There was more particulate organic matter within and between microaggregates in macroaggregates in the presence of earthworms. The larger amounts of organic matter inside stable microaggregates in casts than in bulk soil after 12 days of incubation (140 mg ¹³C kg^?1 soil compared with 20 mg ¹³C kg^?1 soil) indicates that these microaggregates are formed rapidly around freshly incorporated residues within casts. In conclusion, earthworms have a direct impact on the formation of stable microaggregates and the incorporation of organic matter inside these microaggregates, and it seems likely that their activity is of great significance for the long‐term stabilization of organic matter in soils.     

Earthworm populations in relation to soil organic matter dynamics and management in California tomato cropping systems

Earthworms are key regulators of soil structure and soil organic matter (SOM) dynamics in many agroecosystems. They are greatly impacted by agricultural management, yet little is known about how these factors interact to control SOM dynamics. This study sought to explore linkages between agricultural management, earthworms and aggregate associated SOM dynamics through a survey of tomato (Solanum lycopersicum L.) cropping systems in northern California. Earthworms and soil samples were collected between February and April of 2005 from 16 fields under one of three types of residue management: (1) tomato mulch – no postharvest tillage and tomato residues left on the soil surface, (2) cover crop – tomato residues tilled in and leguminous cover crop planted, and (3) bare fallow – tomato residues tilled in and soil surface left exposed throughout the winter. Earthworms were collected via hand-sorting and identified to species, while soils were wet sieved to yield four aggregate size classes: large macroaggregates (>2000 μm), small macroaggregates (250–2000 μm), microaggregates (53–250 μm) and the silt and clay fraction (<53 μm). The combined large and small macroaggregate fraction was then fractionated into coarse particulate organic matter (cPOM; 250 μm), microaggregates within macroaggregates (mM; 53–250 μm) and macroaggregate occluded silt and clay (Msc; <53 μm). The earthworms identified in this survey were composed entirely of exotic species and were dominated by Aporrectodea caliginosa. Earthworm abundance was related to residue management, with the tomato mulch systems averaging 4.5 times greater fresh earthworm biomass than bare fallow (P = 0.024). Aggregate stability and total soil C and N also appeared to be influenced by residue management, such that the tomato mulch system displayed significantly greater mean weight diameters than the bare fallow system (P = 0.049), as well as more than 50% greater total soil C and N (P = 0.049 and P = 0.036; respectively). Earthworm biomass was also found to be positively correlated with total soil C (P = 0.009, R² = 0.39) and N (P = 0.010, R² = 0.039) as well as the proportion of macroaggregate C in the cPOM fraction (P = 0.028, R² = 0.30). Our findings suggest that residue handling and the associated management practices (e.g., tillage, organic vs. conventional agriculture) are important for both earthworm populations and SOM storage. Although earthworms are known to influence SOM in many ways, other factors appear to play a more prominent role in governing aggregate associated SOM dynamics.     

Earthworm impacts on soil organic matter and fertilizer dynamics in tropical hillside agroecosystems of Honduras

Earthworms are important processors of soil organic matter (SOM) and nutrient turnover in terrestrial ecosystems. In agroecosystems, they are often seen as beneficial organisms to crop growth and are actively promoted by farmers and extension agents, yet their contribution to agroecosystem services is uncertain and depends largely on management. The Quesungual slash-and-mulch agroforestry system (QSMAS) of western Honduras has been proposed as a viable alternative to traditional slash-and-burn (SB) practices and has been shown to increase earthworm populations, yet the effect of earthworms on soil fertility and SOM in QSMAS is poorly understood. This study examined the role of Pontoscolex corethrurus in QSMAS by comparing their influence on aggregate-associated SOM and fertilizer dynamics with their effects under SB and secondary forest in a replicated field trial. Both the fertilized QSMAS and SB treatments had plots receiving additions of inorganic ¹⁵N and P, as well as plots with no inorganic N additions. Earthworm populations were manipulated in field microcosms at the beginning of the rainy season within each management treatment via additions of P. corethrurus or complete removal of existing earthworm populations. Microcosms were destructively sampled at harvest of Zea mays and soils were wet-sieved (using 53, 250 and 2000 μm mesh sizes) to isolate different aggregate size fractions, which were analyzed for total C, N and ¹⁵N. The effects of management system were smaller than expected, likely due to disturbance associated with the microcosm installation. Contrary to our hypothesis that earthworms would stabilize organic matter in soil aggregates, P. corethrurus decreased total soil C by 3% in the surface layer (0-15 cm), predominantly through a decrease in the C concentration of macroaggregates (>250 μm) and a corresponding depletion of C in coarse particulate organic matter occluded within macroaggregates. Earthworms also decreased bulk density by over 4%, but had no effect on aggregate size distribution. Within the two fertilized treatments, the QSMAS appeared to retain slightly more fertilizer derived N in smaller aggregate fractions (<250 μm) than did SB, while earthworms greatly reduced the recovery of fertilizer N (34% decrease) in both systems. Although management system did not appear to influence the impact of P. corethrurus on SOM or nutrient dynamics, we suggest the lack of differences may be due to artificially low inputs of fresh residue C to microcosms within all management treatments. Our findings highlight the potential for P. corethrurus to have deleterious impacts on soil C and fertilizer N dynamics, and emphasize the need to fully consider the activities of soil fauna when evaluating agroecosystem management options.     

Decomposition of N-labelled maize leaves in soil affected by endogeic geophagous Aporrectodea caliginosa

A microcosm experiment was carried out for 56 days at 12 °C to evaluate the feeding effects of the endogeic geophagous earthworm species Aporrectodea caliginosa on the microbial use of ¹⁵N-labelled maize leaves (Zea mays) added as 5 mm particles equivalent to 1 mg C and 57 μg N g⁻¹ soil. The dry weight of A. caliginosa biomass decreased in the no-maize treatment by 10% during the incubation and increased in the maize leaf treatments by 18%. Roughly 5% and 10% of the added maize leaf-C and leaf-N, respectively, were incorporated into the biomass of A. caliginosa. About 29% and 33% of the added maize leaf-C were mineralised to CO₂ in the no-earthworm and earthworm treatments, respectively. The presence of A. caliginosa significantly increased soil-derived CO₂ production by 90 μg g⁻¹ soil in the no-maize and maize leaf treatments, but increased the maize-derived CO₂ production only by 40 μg g⁻¹ soil. About 10.5% of maize leaf-C and leaf-N was incorporated into the soil microbial biomass in the absence of earthworms, but only 6% of the maize leaf-C and 3% of the maize leaf-N in the presence of earthworms. A. caliginosa preferentially fed on N rich, maize leaf-colonizing microorganisms to meet its N demand. This led to a significantly increased C/N ratio of the unconsumed microbial biomass in soil. The ergosterol-to-microbial biomass C ratio was not significantly decreased by the presence of earthworms. A. caliginosa did not directly contribute to comminution of plant residues, as indicated by the absence of any effects on the contents of the different particulate organic matter fractions, but mainly to grazing of residue-colonizing microorganisms, increasing their turnover considerably.     

Earthworms bring compacted and loose soil to a similar mechanical state

We tested the hypothesis that earthworms stabilise loose soil and loosen compacted soil to a similar mechanical state. Casts collected from initially loose soil (980 kg m⁻³) had 10-fold greater viscosity (31 kPa s) and 5-fold greater yield stress (200 Pa) than a control soil without worms. Lumbricus terrestris; Dendrobaena sp. and Aporrectodea longa were all investigated, with no difference found between species. In compacted soils (1300 kg m⁻³), A. longa produced casts with similar mechanical properties to loose soil, with viscosity and yield stress decreased by 45% compared to the control without worms. Earthworms were shown to bring initially loose and compacted soil to an intermediate mechanical state that is more favourable for structural stability and root growth.     

Elevated CO2 concentration and nitrogen fertilisation effects on N2O and CH4 fluxes and biomass production of Phleum pratense on farmed peat soil

The effects of elevated CO₂ supply on N₂O and CH₄ fluxes and biomass production of Phleum pratense were studied in a greenhouse experiment. Three sets of 12 farmed peat soil mesocosms (10 cm dia, 47 cm long) sown with P. pratense and equally distributed in four thermo-controlled greenhouses were fertilised with a commercial fertiliser in order to add 2, 6 or 10 g N m⁻². In two of the greenhouses, CO₂ concentration was kept at atmospheric concentration (360 μmol mol⁻¹) and in the other two at doubled concentration (720 μmol mol⁻¹). Soil temperature was kept at 15 °C and air temperature at 20 °C. Natural lighting was supported by artificial light and deionized water was used to regulate soil moisture. Forage was harvested and the plants fertilised three times during the basic experiment, followed by an extra fertilisations and harvests. At the end of the experiment CH₄ production and CH₄ oxidation potentials were determined; roots were collected and the biomass was determined. From the three first harvests the amount of total N in the aboveground biomass was determined. N₂O and CH₄ exchange was monitored using a closed chamber technique and a gas chromatograph. The highest N₂O fluxes (on average, 255 μg N₂O m⁻² h⁻¹ during period IV) occurred just after fertilisation at high water contents, and especially at the beginning of the growing season (on average, 490 μg N₂O m⁻² h⁻¹ during period I) when the competition of vegetation for N was low. CH₄ fluxes were negligible throughout the experiment, and for all treatments the production and oxidation potentials of CH₄ were inconsequential. Especially at the highest rates of fertilisation, the elevated supply of CO₂ increased above- and below-ground biomass production, but both at the highest and lowest rates of fertilisation, decreased the total amount of N in the aboveground dry biomass. N₂O fluxes tended to be higher under doubled CO₂ concentrations, indicating that increasing atmospheric CO₂ concentration may affect N and C dynamics in farmed peat soil.     

Chemical composition, or quality, of agroforestry residues influences N2O emissions after their addition to soil

Emissions of N₂O were measured following addition of ¹⁵N-labelled (2.6-4.7 atom% excess ¹⁵N) agroforestry residues (Sesbania sesban, mixed Sesbania/Macroptilium atropurpureum, Crotalaria grahamiana and Calliandra calothyrsus) to a Kenyan oxisol at a rate of 100 mg N kg soil⁻¹ under controlled environment conditions. Emissions were increased following addition of residues, with 22.6 mg N m⁻² (124.4 mg N m⁻² kg biomass⁻¹; 1.1 mg ¹⁵N m⁻²; 1.03% of ¹⁵N applied) emitted as N₂O over 29 d after addition of both Sesbania and Macroptilium residues in the mixed treatment. Fluxes of N₂O were positively correlated with CO₂ fluxes, and N₂O emissions and available soil N were negatively correlated with residue lignin content (r=−0.49;P<0.05), polyphenol content (r=−0.94;P<0.05), protein binding capacity (r=−0.92;P<0.05) and with (lignin+polyphenol)-to-N ratio (r=−0.55;P<0.05). Lower emission (13.6 mg N m⁻² over 29 d; 94.5 mg N m⁻² kg biomass⁻¹; 0.6 mg ¹⁵N m⁻²; 0.29% of ¹⁵N applied) after addition of Calliandra residue was attributed to the high polyphenol content (7.4%) and high polyphenol protein binding capacity (383 μg BSA mg plant⁻¹) of this residue binding to plant protein and reducing its availability for microbial attack, despite the residue having a N content of 2.9%. Our results indicate that residue chemical composition, or quality, needs to be considered when proposing mitigation strategies to reduce N₂O emissions from systems relying on incorporation of plant biomass, e.g. improved-fallow agroforestry systems, and that this consideration should extend beyond the C-to-N ratio of the residue to include polyphenol content and their protein binding capacity.     

Increased N deposition retards mineralization of old soil organic matter

The aim of this study was to investigate the effects of increased N deposition on new and old pools of soil organic matter (SOM). We made use of a 4-yr experiment, where spruce and beech growing on an acidic loam and a calcareous sand were exposed to increased N deposition (7 vs. 70 kg N ha⁻¹ yr⁻¹) and to elevated atmospheric CO₂. The added CO₂ was depleted in ¹³C, which enabled us to distinguish between old and new C in SOM-pools fractionated into particle sizes. Elevated N deposition for 4 yr increased significantly the contents of total SOM in 0-10 cm depth of the acidic loam (+9%), but not in the calcareous sand. Down to 25 cm soil depth, C storage in the acidic loam was between 100 and 300 g C m⁻² larger under high than under low N additions. However, this increase was small as compared with the SOM losses of 600-700 g C g C 0.25 m⁻¹ m⁻² from the calcareous sand resulting from the disturbance of soils during setting up of the experiment. The amounts of new, less than 4 yr old SOM in the sand fractions of both soils were greater under high N deposition, showing that C inputs from trees into soils increased. Root biomass in the acidic loam was larger under N additions (+25%). Contents of old, more than 4 yr old C in the clay and silt fractions of both soils were significantly greater under high than under low N deposition. Since clay- and silt-bound SOM consists of humified compounds, this indicates that N additions retarded mineralization of old and humified SOM. The retardation of C mineralization in the clay and silt fraction accounted for 60-80 g C m⁻² 4 yr⁻¹, which corresponds to about 40% of the old SOM mineralized in these fraction. As a consequence, preservation of old and humified SOM under elevated N deposition might be a process that could lead to an increased soil C storage in the long-term.     

Performance of para-nitrophenyl phosphate and 4-methylumbelliferyl phosphate as substrate analogues for phosphomonoesterase in soils with different organic matter content

Phosphomonoesterase (PMEase) activity plays a key role in nutrient cycling and is a potential indicator of soil condition and ecosystem stress. We compared para-nitrophenyl phosphate (pNPP) and 4-methylumbelliferyl phosphate (MUP) as substrate analogues for PMEase in 7 natural ecosystem soils and 8 agricultural top soils with contrasting C contents (8.0-414 g kg⁻¹ C) and pH (3.0-7.5). PMEase activities obtained with pNPP (0.05-5 μmol g⁻¹ h⁻¹) were significantly less than activities obtained with MUP (0.9-13 μmol g⁻¹ h⁻¹), especially in soils with a high organic matter content (>130 g kg⁻¹). Only PMEase activities assayed with MUP correlated significantly with total C and total N (r=0.7, P<0.01 all), and pH (r=−0.71, P<0.01). PMEase activities obtained with the two substrate analogues were correlated when expressed on a C-content basis (r=0.8, P<0.001), but not when expressed on an oven-dry soil weight basis. This indicated that interference by organic matter is related to the quantity rather than to the quality of organic matter. Overall, assaying with MUP was more sensitive compared to assaying with pNPP, particularly in the case of high organic and acid soils.     

Earthworm activity as a determinant for N2O emission from crop residue

Earthworm activity may have an effect on nitrous oxide (N₂O) emissions from crop residue. However, the importance of this effect and its main controlling variables are largely unknown. The main objective of this study was to determine under which conditions and to what extent earthworm activity impacts N₂O emissions from grass residue. For this purpose we initiated a 90-day (experiment I) and a 50-day (experiment II) laboratory mesocosm experiment using a Typic Fluvaquent pasture soil with silt loam texture. In all treatments, residue was applied, and emissions of N₂O and carbon dioxide (CO₂) were measured. In experiment I the residue was applied on top of the soil surface and we tested (a) the effects of the anecic earthworm species Aporrectodea longa (Ude) vs. the epigeic species Lumbricus rubellus (Hoffmeister) and (b) interactions between earthworm activity and bulk density (1.06 vs. 1.61 g cm⁻³). In experiment II we tested the effect of L. rubellus after residue was artificially incorporated in the soil. In experiment I, N₂O emissions in the presence of earthworms significantly increased from 55.7 to 789.1 μg N₂O-N kg⁻¹ soil (L. rubellus; p<0.001) or to 227.2 μg N₂O-N kg⁻¹ soil (A. longa; p<0.05). This effect was not dependent on bulk density. However, if the residue was incorporated into the soil (experiment II) the earthworm effect disappeared and emissions were higher (1064.2 μg N₂O-N kg⁻¹ soil). At the end of the experiment and after removal of earthworms, a drying/wetting and freezing/thawing cycle resulted in significantly higher emissions of N₂O and CO₂ from soil with prior presence of L. rubellus. Soil with prior presence of L. rubellus also had higher potential denitrification. We conclude that the main effect of earthworm activity on N₂O emissions is through mixing residue into the soil, switching residue decomposition from an aerobic and low denitrification pathway to one with significant denitrification and N₂O production. Furthermore, A. longa activity resulted in more stable soil organic matter than L. rubellus.     

Use of 13C and 15N natural abundance techniques to characterize carbon and nitrogen dynamics in composting and in compost-amended soils

Isotope fractionation during composting may produce organic materials with a more homogenous δ¹³C and δ¹⁵N signature allowing study of their fate in soil. To verify this, C, N, δ¹³C and δ¹⁵N content were monitored during nine months covered (thermophilic; >40 °C) composting of corn silage (CSC). The C concentration reduced from 10.34 to 1.73 g C (g ash)⁻¹, or 83.3%, during composting. Nitrogen losses comprised 28.4% of initial N content. Compost δ¹³C values became slightly depleted and increasingly uniform (from −12.8±0.6‰ to −14.1±0.0‰) with composting. Compost δ¹⁵N values (0.3±1.3 to 8.2±0.4‰) increased with a similar reduced isotope variability.The fate of C and N of diverse composts in soil was subsequently examined. C, N, δ¹³C, δ¹⁵N content of whole soil (0-5 cm), light (<1.7 g cm⁻³) and heavy (>1.7 g cm⁻³) fraction, and (250-2000 μm; 53-250 μm and <53 μm) size separates, were characterized. Measurements took place one and two years following surface application of CSC, dairy manure compost (DMC), sewage sludge compost (SSLC), and liquid dairy manure (DM) to a temperate (C₃) grassland soil. The δ¹³C values and total C applied (Mg C ha⁻¹) were DM (−27.3‰; 2.9); DMC (−26.6‰; 10.0); SSLC (−25.9‰; 10.9) and CSC (−14.0‰; 4.6 and 9.2). The δ¹³C of un-amended soil exhibited low spatial (−28.0‰±0.2; n=96) and temporal (±0.1‰) variability. All C₄ (CSC) and C₃ (DMC; SSLC) composts, except C₃ manure (DM), significantly modified bulk soil δ¹³C and δ¹⁵N. Estimates of retention of compost C in soil by carbon balance were less sensitive than those calculated by C isotope techniques. One and two years after application, 95 and 89% (CSC), 75 and 63% (SSLC) and 88 and 42% (DMC) of applied compost C remained in the soil, with the majority (80-90%) found in particulate (>53 μm) and light fractions. However, C₄ compost (CSC) was readily detectable (12% of compost C remaining) in mineral (<53 μm) fractions. The δ¹⁵N-enriched N of compost supported interpretation of δ¹³C data. We can conclude that composts are highly recalcitrant with prolonged C storage in non-mineral soil fractions. The sensitivity of the natural abundance tracer technique to characterize their fate in soil improves during composting, as a more homogeneous C isotope signature develops, in addition to the relatively large amounts of stable C applied in composts.     

Organic matter turnover in soil physical fractions following woody plant invasion of grassland: Evidence from natural C and N

Soil physical structure causes differential accessibility of soil organic carbon (SOC) to decomposer organisms and is an important determinant of SOC storage and turnover. Techniques for physical fractionation of soil organic matter in conjunction with isotopic analyses (δ¹³C, δ¹⁵N) of those soil fractions have been used previously to (a) determine where organic C is stored relative to aggregate structure, (b) identify sources of SOC, (c) quantify turnover rates of SOC in specific soil fractions, and (d) evaluate organic matter quality. We used these two complementary approaches to characterize soil C storage and dynamics in the Rio Grande Plains of southern Texas where C₃ trees/shrubs (δ¹³C=−27‰) have largely replaced C₄ grasslands (δ¹³C=−14‰) over the past 100-200 years. Using a chronosequence approach, soils were collected from remnant grasslands (Time 0) and from woody plant stands ranging in age from 10 to 130 years. We separated soil organic matter into specific size/density fractions and determined their C and N concentrations and natural δ¹³C and δ¹⁵N values. Mean residence times (MRTs) of soil fractions were calculated based on changes in their δ¹³C with time after woody encroachment. The shortest MRTs (average=30 years) were associated with all particulate organic matter (POM) fractions not protected within aggregates. Fine POM (53-250 μm) within macro- and microaggregates was relatively more protected from decay, with an average MRT of 60 years. All silt+clay fractions had the longest MRTs (average=360 years) regardless of whether they were found inside or outside of aggregate structure. δ¹⁵N values of soil physical fractions were positively correlated with MRTs of the same fractions, suggesting that higher δ¹⁵N values reflect an increased degree of humification. Increased soil C and N pools in wooded areas were due to both the retention of older C₄-derived organic matter by protection within microaggregates and association with silt+clay, and the accumulation of new C₃-derived organic matter in macroaggregates and POM fractions.     

Interactive effects of functionally different earthworm species on aggregation and incorporation and decomposition of newly added residue carbon

The interactive effects of two functionally different earthworm species (Aporrectodea caliginosa (endogeic species) and Lumbricus rubellus (epigeic species)) on the incorporation of fresh residue into large macroaggregates and formation of microaggregates within these large macroaggregates were investigated during a short-term laboratory experiment using ¹³C-labelled sorghum (Sorghum bicolor (L.) Moench) residues. Soil was collected from a long-term no-tillage agricultural field, crushed through a 250-μm sieve and incubated under laboratory conditions. The following earthworm treatments were applied: (i) soil+¹³C-labelled residue+A. caliginosa; (ii) soil+¹³C-labelled residue+L. rubellus; (iii) soil+¹³C-labelled residue+A. caliginosa+L. rubellus and; (iv) soil+¹³C-labelled residue. Two residue placement treatments (i.e. surface and incorporated) were superimposed on the earthworm treatments. Earthworms were added after 8 days of incubation. Aggregate size distribution and total C and ¹³C were measured after 22 days. Microaggregates, fine inter-microaggregate particulate organic matter (inter-POM) and intra-microaggregate POM (intra-POM) were isolated from macroaggregates. Earthworms had a greater stimulating effect on the formation of large macroaggregates (>2000 μm) and microaggregates within large macroaggregates when residue was incorporated in the soil, especially in the presence of A. caliginosa. When residue was placed on the surface, residue-derived intra-POM C was highest when L. rubellus was present and significantly lower in the presence of A. caliginosa. Residue-derived inter-POM C was highest when a mix of both species was present. These results indicate that earthworm species differentially affect incorporation of fresh organic matter into stable microaggregates within macroaggregates, and that interactive effects of earthworm species might have important consequences for the incorporation and protection of C inside of microaggregates within macroaggregates especially when residues are placed on the soil surface.     

Competition between invasive earthworms (Amynthas corticis, Megascolecidae) and native North American millipedes (Pseudopolydesmus erasus, Polydesmidae): Effects on carbon cycling and soil structure

Invasive earthworms can have significant impacts on C dynamics through their feeding, burrowing, and casting activities, including the protection of C in microaggregates and alteration of soil respiration. European earthworm invasion is known to affect soil micro- and mesofauna, but little is known about impacts of invasive earthworms on other soil macrofauna. Asian earthworms (Amynthas spp.) are increasingly being reported in the southern Appalachian Mountains in southeastern North America. This region is home to a diverse assemblage of native millipedes, many of which share niches with earthworm species. This situation indicates potential for earthworm-millipede competition in areas subject to Amynthas invasion.In a laboratory microcosm experiment, we used two ¹³C enriched food sources (red oak, Quercus rubra, and eastern hemlock, Tsuga canadensis) to assess food preferences of millipedes (Pseudopolydesmus erasus), to determine the effects of millipedes and earthworms (Amynthas corticis) on soil structure, and to ascertain the nature and extent of the interactions between earthworms and millipedes. Millipedes consumed both litter species and preferred red oak litter over eastern hemlock litter. Mortality and growth of millipedes were not affected by earthworm presence during the course of the experiment, but millipedes assimilated much less litter-derived C when earthworms were present.Fauna and litter treatments had significant effects on soil respiration. Millipedes alone reduced CO₂ efflux from microcosms relative to no fauna controls, whereas earthworms alone and together with millipedes increased respiration, relative to the no fauna treatment. CO₂ derived from fresh litter was repressed by the presence of macrofauna. The presence of red oak litter increased CO₂ efflux considerably, compared to hemlock litter treatments.Millipedes, earthworms, and both together reduced particulate organic matter. Additionally, earthworms created significant shifts in soil aggregates from the 2000-250 and 250-53 μm fractions to the >2000 μm size class. Earthworm-induced soil aggregation was lessened in the 0-2 cm layer in the presence of millipedes. Earthworms translocated litter-derived C to soil throughout the microcosm.Our results suggest that invasion of ecosystems by A. corticis in the southern Appalachian Mountains is unlikely to be limited by litter species and these earthworms are likely to compete directly for food resources with native millipedes. Widespread invasion could cause a net loss of C due to increased respiration rates, but this may be offset by C protected in water-stable soil aggregates.