Organic carbon sequestration in earthworm burrows

Earthworms strongly affect soil organic carbon cycling. The aim of this study was to determine whether deep burrowing anecic earthworms enhance carbon storage in soils and decrease C turnover. Earthworm burrow linings were separated into thin cylindrical sections with different distances from the burrow wall to determine gradients from the burrow wall to the surrounding soil. Organic C, total N, radiocarbon (¹⁴C) concentration, stable isotope values (δ¹³C, δ¹⁵N) and extracellular enzyme activities were measured in these samples. Anecic earthworms increased C stocks by 270 and 310 g m^?2 accumulated in the vertical burrows. C-enrichment of the burrow linings was spatially highly variable within a distance of millimetres around the burrow walls. It was shown that C accumulation in burrows can be fast with C sequestration rates of about 22 g C m^?2 yr^?1 in the burrow linings, but accumulated C in the burrows may be mineralised fast with turnover times of only 3–5 years. Carbon stocks in earthworm burrows strongly depended on the earthworm activity which maintains continuous C input into the burrows. The enhanced extracellular enzyme activity of fresh casts was not persistent, but was 47% lower in inhabited burrows and 62% lower in abandoned burrows. Enzyme activities followed the C concentrations in the burrows and were not further suppressed due to earthworms. Radiocarbon concentrations and stable isotopes in the burrow linings showed an exponential gradient with the youngest and less degraded organic matter in the innermost part of the burrow wall. Carbon accumulation by anecic earthworm is restricted to distinct burrows with less influence to the surrounding soil. Contrary to the initial hypothesis, that organic C is stabilised due to earthworms, relaxation time experiments with nuclear magnetic resonance spectroscopy (NMR) did not reveal any enhanced adsorption of C on iron oxides with C stabilising effect. Our results suggest that earthworm activity does not substantially increase subsoil C stocks but burrows serve as fast ways for fresh C transport into deep soil horizons.     

Relationships between earthworm communities and burrow numbers under different land use systems

This study addresses the influence of three different land use systems (continuous maize, pasture/maize rotation, permanent pasture) on the relationships between earthworm populations and the number of earthworm burrows quantified in a soil profile. Quantified burrows were limited to those observable by the naked eye (i.e. >2 mm in diameter) and enumerated earthworms were limited to those which could have created the observable burrows (i.e. >0.3 g).The results were combined with data from the literature coming from different geographical regions. This study showed that earthworm abundance decreased with the increasing land management intensity (maize crop vs. pasture), while the number of burrows could be higher or similar under maize compared to pasture. Under maize, despite lower earthworm abundance and the annual destruction of the burrows by tillage, the number of burrows was almost as high as under pasture. This absence of a relationship between burrow numbers and earthworm abundance was observed in the soil profile and for each layer of the profile for each land use system. Furthermore, the burrow number/m^?2 per earthworm strongly varied depending on land use and was far higher under maize when compared to pasture (74 vs. 7). Therefore, a power-law type relationship was clearly established between burrow number/m^?2 per earthworm and earthworm abundance. This power type relationship was also observed when including data from the literature although it followed a different mathematical model. These results were explained by (i) increased earthworm burrowing activity (i.e. an increase in the number of burrows produced by each earthworm) under maize, and (ii) the dynamics of burrow number under pasture (i.e. decreased burrowing activity and burrow destruction process); both results of food accessibility combined with inter-individual competition. The results of the study suggest that farmers should not use the number of pores as an indicator of earthworm abundance, but as an indicator of earthworm activity, which could be integrated in an indicator of soil quality.     

Impact of the antibiotic sulfadiazine and pig manure on the microbial community structure in agricultural soils

Large amounts of veterinary antibiotics enter soil via manure of treated animals. The effects on soil microbial community structure are not well investigated. In particular, the impact of antibiotics in the presence of manure is poorly understood. In this study, two agricultural soils, a sandy Cambisol (KS) and a loamy Luvisol (ML), were spiked with manure and sulfadiazine (SDZ; 0, 10 and 100 μg g^?1) and incubated for 1, 4, 32 and 61 days. Untreated controls received only water. The microbial community structure was characterised by investigating phospholipid fatty acids (PLFA) and using PCR–denaturing gradient gel electrophoresis (DGGE) of 16S rDNA. The total concentration of PLFA increased with addition of manure and was reduced by both SDZ concentrations at incubation times >4 days. The SDZ addition decreased the bacteria:fungi ratio. The largest stress level, measured as ratio of PLFA (cyc17:0 + cyc19:0)/(16:1ω7c + 18:1ω7c), was found for the controls, followed by the manure treatments and the SDZ treatments. A discriminant analysis of the PLFA clearly separated treatments and incubation times. Both soils differed in total PLFA concentrations and Gram^?:Gram⁺ ratios, but showed similar changes in PLFA pattern upon soil treatment. Effects of manure and SDZ on the bacterial community structure were also revealed by DGGE analysis. Effects on pseudomonads and β-proteobacteria were less pronounced. While community structure remained altered even after two months, the extractable concentrations of SDZ decreased exponentially and the remaining solution concentrations after 32 days were ≤27% of the spiking concentration. Our results demonstrate that a single addition of SDZ has prolonged effects on the microbial community structure in soils.     

Burrow systems of endogeic earthworms: Effects of earthworm abundance and consequences for soil water infiltration

By creating burrows, earthworms influence the transfer properties of soils. The effects of endogeic species on soil transfer properties, however, are not yet well understood because these earthworms generally create burrows that are refilled by casts and have no preferential vertical orientation. Thirty soil cores were incubated for various periods (1–3 or 4 weeks) at different earthworm densities (70, 210, 345 or 480 individuals m⁻²). The cores were then scanned using X-ray tomography and the burrow systems were characterised by measuring the total burrow volume, bioturbation volume (refilled burrows and lateral compaction around the burrows), the number of branches, tortuosity and continuity (assessed by computing the number of burrows with a vertical extension greater than 15, 20 and 25% of the core). We also computed the mean geodesic distance, i.e. the mean distance from the bottom to the top of the core assuming that distances inside burrows are null. Rainfall simulations were carried out on 17 cores chosen to encompass the variations observed in the burrow systems. The water transfer efficiency of each core was estimated by measuring two parameters: breakthrough volume and the percentage of water transmitted after 1 h of rain. Burrow and bioturbation volume increased significantly and steadily with time and earthworm density. We estimated that on average Allolobophora chlorotica burrowed 22 cm per week. All other burrow system characteristics also increased with time and earthworm density except the mean geodesic distance, which decreased significantly. This suggests that intraspecific interactions had no significant effect on burrow system geometry. Univariate PLS regressions were used to understand which burrow system characteristics had the strongest influence on water transfer. These regressions showed that the mean geodesic distance was the most important parameter. This means that in addition to individual burrow characteristics, the spatial arrangement of the whole burrow system also had a major effect on transfer properties.     

What are the limits of the drilosphere? An incubation experiment using Metaphire posthuma

The near infrared reflectance spectroscopy (NIRS) method was used in the present study to compare earthworm-made soil aggregates to aggregates found in the surrounding bulk soil. After initially assessing the daily cast production of Metaphire posthuma, boxes with soil incubated with M. posthuma and control soils were subjected to wetting in order to reorganize the soil structure. After two months of incubation, soil aggregates produced by earthworms (casts and burrows), soil aggregates that were appeared to be unaffected by earthworms (bulk soil without visible trace of earthworm bioturbation from the earthworm treatment) and soil aggregates that were entirely unaffected by earthworms (control – no earthworm – treatment) were sampled and their chemical signatures analyzed by NIRS. The production of below-ground and surface casts reached 14.9 g soil g worm^?1 d^?1 and 1.4 g soil g worm^?1 d^?1, respectively. Soil aggregates from the control soils had a significantly different NIRS signature from those sampled from boxes with earthworms. However, within the earthworm incubation boxes the NIRS signature was similar between cast and burrow aggregates and soil aggregates from the surrounding bulk soil. We conclude that the high cast production by M. posthuma and the regular reorganization of the soil structure by water flow in and through the soil lead to a relatively homogenous soil structure. Given these results, we question the relevance of considering the bulk soil that has no visible activity of earthworm activity as a control to determine the effect of earthworms on soil functioning.     

Quantitative estimates of burrow construction and destruction,by anecic and endogeic earthworms in repacked soil cores

Although the role of earthworms in soil functioning is often emphasised, many important aspects of earthworm behaviour are still poorly understood. In this study we propose a simple and cost-effective method for estimating burrow system area and continuity, as well as a new and often neglected parameter, the percentage of burrow refilling by the earthworms own casts. This novel parameter is likely to have a huge influence on the transfer properties of the burrow system. The method uses standard repacked soil cores in PVC cylinders and takes advantages of clay shrinkage and the fact that earthworms were previously shown to prefer to burrow at the PVC/soil interface. In this way, after removing the PVC cylinders off dry cores, the external section of the burrow system made by earthworms along the soil walls could be easily described. We applied this method to characterise the burrow systems of four earthworms species: two anecics (Aporrectodea caliginosa nocturna and Aporrectodea caliginosa meridionalis) and two endogeics (Aporrectodea caliginosa icaliginosa and Allolobophora chlorotica). After one month the burrow's area generated by both anecic species were much larger (about 40 cm²) than the endogeic burrow's area (about 15 cm²). A. nocturna burrow system continuity was higher than that of A. meridionalis and both anecic burrow systems were more continuous than those made by the endogeic earthworms. This was partly explained by the far larger proportion of the burrow area that was refilled with casts: approximately 40% and 50% for Al. chlorotica and A. caliginosa, respectively compared with approximately 20% for the anecic burrows. We discuss whether these estimates could be used in future models simulating the dynamics of earthworm burrow systems by taking into account both burrow creation and destruction by earthworms.     

Distinct microbial and faunal communities and translocated carbon in Lumbricus terrestris drilospheres

Lumbricus terrestris is a deep-burrowing anecic earthworm that builds permanent, vertical burrows with linings (e.g., drilosphere) that are stable and long-lived microhabitats for bacteria, fungi, micro- and mesofauna. We conducted the first non-culture based field study to assess simultaneously the drilosphere (here sampled as 0–2 mm burrow lining) composition of microbial and micro/mesofaunal communities relative to bulk soil. Our study also included a treatment of surface-applied ¹³C- and ¹⁵N-labeled plant residue to trace the short-term (40 d) translocation of residue C and N into the drilosphere, and potentially the assimilation of residue C into drilosphere microbial phospholipid fatty acids (PLFAs). Total C concentration was 23%, microbial PLFA biomass was 58%, and PLFAs associated with protozoa, nematodes, Collembola and other fauna were 200-to-300% greater in the drilosphere than in nearby bulk soil. Principal components analysis of community PLFAs revealed that distributions of Gram-negative bacteria and actinomycetes and other Gram-positive bacteria were highly variable among drilosphere samples, and that drilosphere communities were distinct from bulk soil communities due to the atypical distribution of PLFA biomarkers for micro- and mesofauna. The degree of microbial PLFA ¹³C enrichment in drilosphere soils receiving ¹³C-labeled residue was highly variable, and only one PLFA, 18:1ω9c, was significantly enriched. In contrast, 11 PLFAs from diverse microbial groups where enriched in response to residue amendment in bulk soil 0–5 cm deep. Among control soils, however, a significant δ¹³C shift between drilosphere and bulk soil at the same depth (5–15 cm) revealed the importance of L. terrestris for translocating perennial ryegrass-derived C into the soil at depth, where we estimated the contribution of the recent grass C (8 years) to be at least 26% of the drilosphere soil C. We conclude that L. terrestris facilitates the translocation of plant C into soil at depth and promotes the maintenance of distinct soil microbial and faunal communities that are unlike those found in the bulk soil.     

Soil organic matter in soil depth profiles: Distinct carbon preferences of microbial groups during carbon transformation

This study investigates how carbon sources of soil microbial communities vary with soil depth. Microbial phospholipid fatty acids (PLFA) were extracted from 0–20, 20–40 and 40–60 cm depth intervals from agricultural soils and analysed for their stable carbon isotopes (δ¹³C values). The soils had been subjected to a vegetation change from C3 (δ¹³C≈?29.3‰) to C4 plants (δ¹³C≈?12.5‰) 40 years previously, which allowed us to trace the carbon flow from plant-derived input (litter, roots, and root exudates) into microbial PLFA. While bulk soil organic matter (SOM) reflected ≈12% of the C4-derived carbon in top soil (0–20 cm) and 3% in deeper soil (40–60 cm), the PLFA had a much higher contribution of C4 carbon of about 64% in 0–20 cm and 34% in 40–60 cm. This implies a much faster turnover time of carbon in the microbial biomass compared to bulk SOM. The isotopic signature of bulk SOM and PLFA from C4 cultivated soil decreases with increasing soil depth (?23.7‰ to ?25.0‰ for bulk SOM and ?18.3‰ to ?23.3‰ for PLFA), which demonstrates decreasing influence of the isotopic signature of the new C4 vegetation with soil depth. In terms of soil microbial carbon sources this clearly shows a high percentage of C4 labelled and thus young plant carbon as microbial carbon source in topsoils. With increasing soil depth this percentage decreases and SOM is increasingly used as microbial carbon source. Among all PLFA that were associated to different microbial groups it could be observed that (a) depended on availability, Gram-negative and Gram-positive bacteria prefer plant-derived carbon as carbon source, however, (b) Gram-positive bacteria use more SOM-derived carbon sources while Gram-negative bacteria use more plant biomass. This tendency was observed in all three-depth intervals. However, our results also show that microorganisms maintain their preferred carbon sources independent on soil depth with an isotopic shift of 3–4‰ from 0–20 to 40–60 cm soil depth.     

Changes of microbial properties in (near-) rhizosphere soils after phytoextraction by Sedum alfredii H: A rhizobox approach with an artificial Cd-contaminated soil

The ultimate goal of soil remediation is to restore soil health. Soil microbial parameters are considered to be effective indicators of soil health. The aim of this study was to determine the effects of phytoextraction on microbial properties through the measurement of soil microbial biomass carbon, soil basal respiration and enzyme activities. For this purpose, a pre-stratified rhizobox experiment was conducted with the Cd hyperaccumulator Sedum alfredii H. for phytoextraction Cd from an artificial contaminated soil (15.81 mg kg⁻¹) under greenhouse conditions. The plant and soil samples were collected after growing the plant for three and six months with three replications. The results indicated that the ecotype of S. alfredii H. originating from an ancient silver mining site was a Cd-hyperaccumulator as it showed high tolerance to Cd stress, the shoot Cd concentration were as high as 922.6 mg kg⁻¹ and 581.9 mg kg⁻¹ at the two samplings, and it also showed high BF (58.4 and 36.8 after 3 and 6 months growth), and TF (5.8 and 5.1 after 3 and 6 months growth). The amounts of Cd accumulated in the shoots of S. alfredii reached to an average of 1206 μg plant⁻¹ after 6 months growth. Basal respiration, invertase and acid phosphatase activities of the rhizosphere soil separated by the shaking method were significantly higher (P < 0.01) than that of the near-rhizosphere soil and the unplanted soil after 3 months growth, so were microbial biomass carbon, urease, invertase and acid phosphatase activities of the rhizosphere soil after 6 months growth. Acid phosphatase activity of the 0–2 mm sub-layer rhizosphere soil collected by the pre-stratified method after 3 months growth was significantly higher (P < 0.05) than that of other sub-layer rhizosphere soils and bulk soil, and so were microbial biomass carbon, basal respiration, urease, invertase and acid phosphatase activities of the 0–2 mm sub-layer rhizosphere soil after 6 months growth. It was concluded that phytoextraction by S. alfredii could improve soil microbial properties, especially in rhizosphere, and this plant poses a great potential for the remediation of Cd contaminated soil.     

Assessment of soil structural differentiation around earthworm burrows by means of X-ray computed tomography and scanning electron microscopy

The burrowing activity of earthworms creates a distinct area around the resulting macropores called the drilosphere, which controls various soil processes. Density and microstructure of the drilosphere were studied and compared with those of the surrounding soil. For this purpose soil cores were separately inoculated with the vertically burrowing earthworm species Lumbricus terrestris. After 70 days some cores were compacted by a hydraulic press (250 kPa) and all cores were analysed by means of X-ray computed tomography. Mean Hounsfield Units were measured for concentric ROI cylinders (ROI = region of interest) of increasing diameters located around vertical macropore sections within selected horizontal slices. Based on these data we estimated stepwise the distribution of bulk density from the inner boundary of the drilosphere to the surrounding soil. In uncompacted soil the bulk density of the drilosphere was increased by 11% over that of the surrounding soil. In cross section, drilosphere and burrow form a concentric area with a total radius up to 2.2 cm. Soil compaction increased the dry bulk density of soil and decreased the diameter of earthworm burrows. Moreover, we found a less dense part of soil between the dense drilosphere and the remaining soil of the compacted core. Scanning electron microscopy revealed that the coarse silt particles of the bulk soil were rearranged to a parallel orientation due to compaction whereas the microstructure of the drilosphere remained unchanged. In any case, the drilosphere revealed a very homogeneous and dense arrangement of silt particles.     

Microbial activity,community structure and potassium dynamics in rhizosphere soil of soybean plants treated with glyphosate

With the advent of glyphosate [N-(phosphonomethyl)glycine] tolerant crops, soils have now been receiving repeated applications of the herbicide for over 10 years in the Midwestern USA. There is evidence that long-term use of glyphosate can cause micronutrient deficiency but little is known about plant potassium (K) uptake interactions with glyphosate. The repeated use of glyphosate may create a selection pressure in soil microbial communities that could affect soil K dynamics and ultimately K availability for crops. Therefore, the objectives of this study were to characterize the effect of foliar glyphosate applied to GR (glyphosate resistant) soybeans on: (1) rhizosphere microbial community profiles using ester linked fatty acid methyl ester (EL-FAME) biomarkers, (2) exchangeable, non-exchangeable, and microbial K in the rhizosphere soil, and (3) concentrations of soybean leaf K. A greenhouse study was conducted in a 2 × 2 × 3 factorial design with two soil treatments (with or without long-term field applications of glyphosate), two plant treatments (presence and absence of soybean plants), and three rates of glyphosate treatments (0×, 1× at 0.87, and 2× at 1.74 kg ae ha^?1, the recommended field rate). After each glyphosate application, rhizosphere soils were sampled and analyzed for microbial community structure using ester linked fatty acid methyl ester biomarkers (EL-FAME), and exchangeable, plant tissue and microbial biomass K. Glyphosate application caused a significant decrease in the total microbial biomass in soybean rhizosphere soil that had no previous exposure to glyphosate, at 7 days after glyphosate application. However, no significant changes were observed in the overall microbial community structure. In conclusion, the glyphosate application lowered the total microbial biomass in the GR soybean rhizosphere soil that had no previous exposure to glyphosate, at 7 days after glyphosate application; caused no changes in the microbial community structure; and did not reduce the plant available K (soil exchangeable or plant tissue K).     

Effect of litter quality and soil fungi on macroaggregate dynamics and associated partitioning of litter carbon and nitrogen

We investigated the effect of plant residue decomposability and fungal biomass on the dynamics of macroaggregate (250–2000 μm) formation in a three months' incubation experiment and determined the distribution of residue-derived C and N in the microbial biomass and in aggregate size fractions (250–2000 μm, 53–250 μm and <53 μm) using ¹³C and ¹⁵N data. A silty loam soil (sieved <250 μm) was incubated with and without addition of ¹⁵N labelled maize leaves (C/N = 27.4) and roots (C/N = 86.4). Each treatment was carried out with and without fungicide application. The addition of maize residues enhanced soil respiration and microbial biomass C and N and resulted in increased macroaggregate formation with a higher and more rapid maximum macroaggregation in the soil amended with maize leaves than in that with addition of roots. Fungicide application led to a significant decline of microbial biomass C and mineralization of the added residues compared to untreated soils, which demonstrates a successful suppression of part of the active microbial biomass by the fungicide. However, this was not confirmed by a generally lower ergosterol concentration. Consequently, ergosterol was no reliable fungal biomarker in periods of rapid decline of the fungal biomass. A single addition of fungicide was insufficient for continued inhibition of the fungal biomass. Yet, a significant delay (28–42 days) in macroaggregation in fungicide treated compared to untreated samples highlighted the importance of the fungal biomass in macroaggregate formation. Macroaggregates were enriched in maize-derived ¹³C and ¹⁵N compared to microaggregates or the fraction < 53 μm. They turned over rapidly with decreasing substrate availability, which entailed a transfer of maize-derived C and N stored within macroaggregates during the first weeks of incubation to microaggregates with proceeding incubation time. Our results indicate that this transfer happened within macroaggregates, because no considerable amount of free particulate organic matter (POM) was released upon macroaggregate breakdown. We conclude that substrate decomposability and fungal activity are key factors determining extent and dynamics of macroaggregation during decomposition processes. Macroaggregate formation implied rapid incorporation and thereby short-term protection of maize-derived C and N. Moreover, macroaggregates allowed a transfer of maize-derived organic matter into microaggregates within macroaggregates, which prevented the release of significant amounts of free POM upon macroaggregate breakdown. Consequently, macroaggregates constitute to the transfer of recently added C into more stable soil organic matter fractions.     

Soil enzyme activities,microbial community composition and function after 47 years of continuous green manuring

Green manuring practices can influence soil microbial community composition and function and there is a need to investigate the influence compared with other types of organic amendment. This study reports long-term effects of green manure amendments on soil microbial properties, based on a field experiment started in 1956. In the experiment, various organic amendments, including green manure, have been applied at a rate of 4 t C ha⁻¹ every second year. Phospholipid fatty acid analysis (PLFA) indicated that the biomass of bacteria, fungi and total microbial biomass, but not arbuscular mycorrhizal (AM) fungi, generally increased due to green manuring compared with soils receiving no organic amendments. Some differences in abundance of different microbial groups were also found compared with other organic amendments (farmyard manure and sawdust) such as a higher fungal biomass and consequently a higher fungal/bacterial ratio compared with amendment with farmyard manure. The microbial community composition (PLFA profile) in the green manure treatment differed from the other treatments, but there was no effect on microbial substrate-utilization potential, determined using the Biolog EcoPlate. Protease and arylsulphatase activities in the green manure treatment were comparable to a mineral fertilized treatment receiving no additional C, whereas acid phosphatase activity increased. It can be concluded that green manuring had a beneficial impact on soil microbial properties, but differed in some aspects to other organic amendments which might be attributed to differences in quality of the amendments.     

Role of soil water content in the carbon and nitrogen dynamics of Lumbricus terrestris L. burrow soil

We evaluated the role of soil water content in controlling C and N dynamics within the drilosphere created by the anecic earthworm Lumbricus terrestris (L.). Mesocosms (volume = 3.1 l) were each amended with corn litter and three earthworms. Control treatments received no earthworms and no other earthworm species were present in the soil. WET and DRY treatments received a total of 9.25 cm and 3.25 cm of water, respectively. Water was added on weeks 1, 3, 7, and 10 at a rate of 2.0 cm per mesocosm for WET treatments and 0.5 cm per mesocosm for DRY treatments. Mesocosms were sampled destructively after incubation at 18–20 °C for 0, 3, 7, and 13 weeks. The water content of WET burrow soil ranged from 0.12 g g⁻¹ to 0.18 g g⁻¹ and was significantly higher than in the DRY treatment throughout the incubation period. The live weight of earthworms was significantly higher in the WET treatment only on week 13, whereas litter consumption was significantly lower in the DRY treatment for week 13. Carbon mineralization, measured as CO₂ evolved after a 24-h incubation, was consistently higher in WET than in DRY burrow soil. Effects of differences in soil water content were also apparent for biomass C and metabolic quotient. Soil water content did no affect the total C concentration of burrow soil. DRY burrow soil had consistently lower levels of nitrate than WET soil throughout the experiment. Lower levels of ammonium and inorganic N were observed for WET burrow soil on weeks 3 and 7. Water content did not have a significant effect on burrow soil total N. We concluded that the water content of the drilosphere affects both C and N dynamics and can affect the speciation of inorganic N; yet, the effects of soil water content do not appear to result from differences in the feeding activities of anecic earthworms.     

Abbau von Isoproturon in Regenwurm‐Makroporen und in der Unterbodenmatrix – Eine Feldstudie

Degradation of isoproturon in earthworm macropores and subsoil matrix—a field study The objective is to compare the time scale of microbial degradation of the herbicide Isoproturon at the end of earthworm burrows with the time scale of microbial degradation in the surrounding soil matrix. To this end, we developed a method which allows the observation of microbial degradation on Isoproturon in macropores under field conditions. Study area was the well‐investigated Weiherbach catchment (Kraichgau, SW Germany). The topsoil of a 12 m² large plot parcel was removed, the parcel was covered with a tent and instrumented with TDR and temperature sensors at two depths. After preliminary investigations to optimize application and sampling techniques, the bottom of 55 earthworm burrows, located at a depth of 80–100 cm, was inoculated with Isoproturon. Within an interval of 8 d, soil material from the bottom of 5–6 earthworm burrows was taken into the laboratory and analyzed for the Isoproturon concentration for investigation of the degradation kinetics. Furthermore, the degradation of Isoproturon in the soil matrix, that surrounded the macropores at the field plot, was observed in the laboratory. Microbial degradation of Isoproturon at the bottom of the earthworm burrows was with a DT‐50‐value of 15.6 d almost as fast as in the topsoil. In the soil matrix that closely surrounded the center of the earthworm burrows, no measurable degradation was observed within 30 d. The clearly slower degradation in the soil matrix may be likely explained by a lower microbial activity that was observed in the surrounding soil matrix. The results give evidence that deterministic modeling of the fate of pesticides once transported into heterogeneous subsoils by preferential flow requires an accuracy of a few centimeters in terms of predicting spatial locations: time scales of microbial degradation in the subsoil drop almost one order of magnitude, in case the herbicides dislocates from the bottom of an earthworm burrow a few centimeter into the surrounding soil matrix. If at all, predictions of such an accuracy can only be achieved at locations at sites where the soil hydraulic properties and the macropore system are known at a very high spatial resolution.     

Microbiological characterization of the structures built by earthworms and ants in an agricultural field

The genesis and architecture of the structures built by ants and earthworms differ markedly, suggesting that—in addition to having different physical and chemical properties—the resident microbial community should also have unique properties. We characterized the inorganic N, biomass C, C mineralization rate, and functional diversity of the microbial communities of earthworm casts, earthworm burrow soil, ant mounds, and bulk soil from an agricultural field. Mound soil was most enriched in inorganic N and had the lowest pH, moisture content, and C mineralization rate. Functional diversity was evaluated by determining the ability of microorganisms to grow on 31 substrates using Biolog^®EcoPlates in combination with a most probable number (MPN) approach. Casts had MPNs that were one to two orders of magnitude higher than burrow, mound and bulk soil for most substrates. Casts also had the highest MPNs for particular substrate guilds relative to bulk soil, followed by mound and burrow soil. Indices of substrate diversity and evenness were highest for casts, followed by burrow, mound, and bulk soil. Differences in the type of habitat provided by the structures built by ants and earthworms result in the differential distribution of nutrients, microbial activity, and metabolic diversity of soils within an agricultural field that affect soil fertility and quality.     

Assessment of earthworm contribution to soil hydrology: a laboratory method to measure water diffusion through burrow walls

The capacity for water diffusion in burrow walls (i.e. the coefficient of sorptivity) either burrowed by Lumbricus terrestris (T-Worm) or artificially created (T-Artificial) was studied through an experimental design in a 2D terrarium. In addition, the soil density of earthworm casts, burrow walls (0–3 mm around the burrow) and the surrounding soil (>3 mm) were measured using the method of petroleum immersion. This study demonstrated that the quantity of water which transits through burrows of L. terrestris in the soil matrix was lower than that transited through soil fractures, due to a reduction of soil porosity in burrow walls (compaction: cast > worms burrow walls > surrounding soil > artificial burrow walls). Earthworm behaviour, in particular burrow reuse with associated cast pressing on walls, could explain the larger burrow wall compaction in earthworm burrows. If water diffusion was lower through the compacted burrows, burrow reuse by the worms makes them more stable (worms would maintain the structure over years) than unused burrows. The present experimental design could be used to test and measure the specific differences between earthworm species in their contributions to water diffusion. Probably, these contributions depend on the presumed related-species behaviours which would determine the degree of burrow wall compaction.     

Effect of soil moisture and bovine urine on microbial stress

While many studies have examined the cycling of urinary nutrients, few have focused on the effects ruminant urine might have on the soil microbial community. Urine application can cause microbial communities to become stressed, potentially changing community composition and microbial function with subsequent effects on nutrient dynamics. Identification of the factors that stress microbes may assist in explaining ruminant urine effects on nutrient cycling. In this laboratory study bovine urine, with either a high (15.0 g K⁺ l^?1) or low (10.4 g K⁺ l^?1) salt concentration, was added to repacked soil cores maintained at high or low soil moisture contents (70 or 35% water-filled pore space, respectively). Control cores did not receive urine. Microbial stress was measured using phospholipid fatty acid (PLFA) biomarker ratios. Urine addition increased stress as indicated by a decrease in the iso15:0/anteiso15:0 PLFA ratio from >1.35 to <0.95 in both wet and dry soils and by an increase in the 18:1ω9trans/18:1ω9cis PLFA ratio from 1.4 to 1.9 from day 8 onwards in wet soils. Higher stress was indicated by a lower Gram-positive/Gram-negative PLFA ratio in the urine treatments than in the control treatments on day 29 and this may have been a response to the reduction in substrate availability as the experiment progressed. The PLFA biomarkers showed that the salt treatments did not induce stress. Stress induced by urine addition and wet soil treatments was also indicated by principal component analyses and the metabolic quotient for CO₂, respectively. Thus microbial stress was induced by both urine addition and high soil moisture content, but not specifically by increasing the urinary salt concentration.     

Influence of microbial populations and residue quality on aggregate stability

Soil structure mediates many biological and physical soil processes and is therefore an important soil property. Physical soil processes, such as aggregation, can be markedly influenced by both residue quality and soil microbial community structure. Three experiments were conducted to examine (i) the temporal dynamics of aggregate formation and the water stability of the obtained aggregates, (ii) the effect of residue quality on aggregation and microbial respiration, and (iii) the effect of fungi and bacteria on aggregation.In the first experiment, 250 μm sieved air-dried soil, mixed with wheat straw, was incubated for 14 days to allow formation of water-stable macroaggregates (>250 μm). Aggregate stability was measured by wet sieving after four different disruptive treatments: (i) soil at field capacity; (ii) soil air-dried and slowly wetted; (iii) soil air-dried and quickly wetted; (iv) 8 mm sieved soil, air-dried and immersed in water (slaking). After 14 days of incubation, maximum aggregation for soil sieved at field capacity was reached; however, these newly formed aggregates were not yet resistant to slaking.During the second experiment, the effect of low-quality residue (C/N: 108) (with or without extra mineral nitrogen) and high-quality residue (C/N: 19.7) (without extra mineral nitrogen) on macroaggregate formation and fungal and bacterial populations was tested. After 14 days, aggregation, microbial respiration, and total microbial biomass were not significantly different between the low-quality (minus mineral nitrogen) and high-quality residue treatment. However, fungal biomass was higher for the low-quality residue treatment compared to the high-quality residue treatment. In contrast, bacterial populations were favored by the high-quality residue treatment. Addition of mineral N in the low-quality residue treatment resulted in reduced macroaggregate formation and fungal biomass, but had no effect on bacterial biomass. These observations are not conclusive for the function of fungal and/or bacterial biomass in relation to macroaggregate formation. In order to directly discern the influence of soil microflora on aggregation, a third experiment was conducted in which a fungicide (captan) or bactericide (oxytetracycline) was applied to selectively suppress fungal or bacterial populations. The direct suppression of fungal growth by addition of fungicide led to reduced macroaggregate formation. However, suppression of bacterial growth by addition of bactericide did not lead to reduced macroaggregate formation. In conclusion, macroaggregate formation was positively influenced by fungal activity but was not significantly influenced by residue quality or bacterial activity.     

Impact of allelochemical exuded from allelopathic rice on soil microbial community

Allelopathic rice releases allelochemicals from its roots to paddy soils at early growth stages to inhibit neighboring weeds. However, little is currently known about the effects of allelochemicals on soil microbes. In this study, we show that allelopathic rice can have great impact on the population and community structure of soil microbes. Allelopathic rice PI312777 seedlings reduced the culturable microbial population and total PLFA when compared to non-allelopathic rice Liaojing-9. Similar results were observed when, instead of growing seedlings, soils were incubated with plant root exudates. This result demonstrates that the composition of root exudates from the rice varieties tested contributes to the soil microbial community. Further experiments showed that the microbial community was affected by the allelochemical 5,4′-dihydroxy-3′,5′-dimethoxy-7-O-β-glucopyranosylflavone exuded from allelopathic rice roots, through immediately hydrolyzing glucose with stimulation on soil bacteria and aglycone (5,7,4′-trihydroxy-3′,5′-dimethoxyflavone) with inhibition on soil fungi. This result indicates that the flavone O-glycoside can provide carbon and interact with soil microbes. PC analysis of the fatty acid data clearly separated the allelopathic PI312777 and the non-allelopathic Liaojing-9 variety (PC1 = 46.4%, PC2 = 20.3%). Similarly, the first principal component (PC1 = 37.4%) together with the second principal component (PC2 = 17.3%) explained 54.7% of the variation between the allelopathic and non-allelopathic root exudates. Furthermore, the canonical correlation between allelopathic root exudates and the flavone O-glycoside was statistically significant (Canonical R = 0.889, χ² (25) = 69.72, p = 0.0041). Although the data generated in this study were not completely consistent between culturable microbes and PLFA profile, it is a fact that variation in soil microbial populations and community structures could be distinguished by the allelopathic and non-allelopathic rice varieties tested. Our results suggest that individual components of rice root exudates, such as allelochemicals from allelopathic rice, can modify the soil microbial community.