Food choice and reproductive success of Folsomia candida feeding on copper-contaminated mycelium of the soil fungus Alternaria alternata

We examined collembolan food preference for fungal mycelium grown on copper-contaminated medium, and the relationship between copper content, food selectivity and collembolan fitness when fed contaminated mycelium.To clarify whether collembolan food selectivity is related to fitness parameters, Folsomia candida were fed mycelium of the dark-pigmented fungus Alternaria alternata grown on medium with different copper concentrations. Copper-contaminated food (fungus grown on 50, 125, 250 and 500 μg Cu g^?1 medium, fresh wt.) was offered together with untreated food for 4 weeks. F. candida fed selectively on the provided mycelium and discriminated clearly between mycelium grown on high and low levels of contamination, distinctly preferring fungus grown on medium with a total copper concentration of 50 and 125 μg g^?1. In contrast, fungus grown on highly contaminated medium (250 and 500 μg g^?1) was avoided. Collembolan food preference generally matched fitness parameters. Reproduction was significantly affected by the total copper concentration of the fungal growth medium. When fed their preferred mycelium, collembolan reproduction was enhanced, whereas a diet of highly contaminated mycelium (250 or 500 μg g^?1) resulted in a strong decrease in reproduction. Adult survival was affected only marginally. Even though heavy metal contamination is a potential stress factor for many soil microarthropods, F. candida is able to discriminate between high and low quality food sources, and even benefits from moderately elevated copper concentrations.     

Emission of nitrous oxide from hydrocarbon contaminated soil amended with waste water sludge and earthworms

Soils in Mexico are often contaminated with hydrocarbons and addition of waste water sludge and earthworms accelerates their removal. However, little is known how contamination and subsequent bioremediation affects emissions of N₂O and CO₂. A laboratory study was done to investigate the effect of waste water sludge and the earthworm Eisenia fetida on emission of N₂O and CO₂ in a sandy loam soil contaminated with the polycyclic aromatic hydrocarbons (PAHs): phenanthrene, anthracene and benzo(a)pyrene. Emissions of N₂O and CO₂, and concentrations of inorganic N (ammonium (NH₄⁺), nitrite (NO₂^?) nitrate (NO₃^?)) were monitored after 0, 5, 24, 72 and 168 h. Adding E. fetida to the PAHs contaminated soil increased CO₂ production rate significantly 2.0 times independent of the addition of sludge. The N₂O emission rate from unamended soil expressed on a daily base was 5 μg N kg^?1 d^?1 for the first 2 h and increased to a maximum of 325 μg N kg^?1 d^?1 after 48 h and then decreased to 10 μg N kg^?1 d^?1 after 168 h. Addition of PAHs, E. fetida or PAHs + E. fetida had no significant effect on the N₂O emission rate. Adding sludge to the soil sharply increased the N₂O emission rate to >400 μg N kg^?1 d^?1 for the entire incubation with a maximum of 1134 μg N kg^?1 d^?1 after 48 h. Addition of E. fetida, PAHs or PAHs + E. fetida to the sludge-amended soil reduced the N₂O emission rate significantly compared to soil amended with sludge after 24 h. It was found that contaminating soil with PAHs and adding earthworms had no effect on emissions of N₂O. Emission of N₂O, however, increased in sludge-amended soil, but addition of earthworms to this soil and contamination reduced it.     

Formation and use of microbial residues after adding sugarcane sucrose to a heated soil devoid of soil organic matter

A 67-day incubation experiment was carried out with a soil initially devoid of any organic matter due to heating, which was amended with sugarcane sucrose (C₄-sucrose with a δ¹³C value of ?10.5‰), inorganic N and an inoculum for recolonisation and subsequently at day 33 with C₃-cellulose (δ¹³C value of ?23.4‰). In this soil, all organic matter is in the microbial biomass or in freshly formed residues, which makes it possible to analyse more clearly the role of microbial residues for decomposition of N-poor substrates. The average δ¹³C value over the whole incubation period was ?10.7‰ in soil total C in the treatments without C₃-cellulose addition. In the CO₂ evolved, the δ¹³C values decreased from ?13.4‰ to ?15.4‰ during incubation. In the microbial biomass, the δ¹³C values increased from ?11.5‰ to ?10.1‰ at days 33 and 38. At day 67, 36% of the C₄-sucrose was left in the treatment without a second amendment. The addition of C₃-cellulose resulted in a further 7% decrease, but 4% of the C₃-cellulose was lost during the second incubation period. Total microbial biomass C declined from 200 μg g^?1 soil at day 5 to 70 μg g^?1 soil at day 67. Fungal ergosterol increased to 1.5 μg g^?1 soil at day 12 and declined more or less linearly to 0.4 μg g^?1 soil at day 67. Bacterial muramic acid declined from a maximum of 35 μg g^?1 soil at day 5 to a constant level of around 16 μg g^?1 soil. Glucosamine showed a peak value at day 12. Galactosamine remained constant throughout the incubation. The fungal C/bacterial C ratio increased more or less linearly from 0.38 at day 5 to 1.1 at day 67 indicating a shift in the microbial community from bacteria to fungi during the incubation. The addition of C₃-cellulose led to a small increase in C₃-derived microbial biomass C, but to a strong increase in C₄-derived microbial biomass C. At days 45 and 67, the addition of N-free C₃-cellulose significantly decreased the C/N ratio of the microbial residues, suggesting that this fraction did not serve as an N-source, but as an energy source.     

Remote estimation of gross primary production in wheat using chlorophyll-related vegetation indices

A number of recent studies have focused on estimating gross primary production (GPP) using vegetation indices (VIs). In this paper, GPP is retrieved as a product of incident light use efficiency (LUE), defined as GPP/PAR, and the photosynthetically active radiation (PAR). As a good correlation is found between canopy chlorophyll content and incident LUE for six types of wheat canopy (R² = 0.87, n = 24), indices aimed for chlorophyll assessment can be used as an indicator of incident LUE and the product of chlorophyll indices and PAR will be a proxy of GPP. In a field experiment, we investigated four canopy chlorophyll content related indices (Red edge Normalized Difference Vegetation Index [Red Edge NDVI], modified Chlorophyll Absorption Ratio Index [MCARI₇₁₀], Red Edge Chlorophyll Index [CI_{red edge}] and the MERIS Terrestrial Chlorophyll Index [MTCI]) for GPP estimation during the growth cycle of wheat. These indices are validated for leaf and canopy chlorophyll estimation with ground truth data of canopy chlorophyll content. With ground truth data, a strong correlation is observed for canopy chlorophyll estimation with correlation coefficients R² of 0.79, 0.84, 0.85 and 0.87 for Red Edge NDVI, MCARI₇₁₀, CI_{red edge} and MTCI, respectively (n = 24). As evidence of the existence of a relationship between canopy chlorophyll and GPP/PAR, these indices are shown to be a good proxy of GPP/PAR with R² ranging from 0.70 for Red Edge NDVI and 0.75 for MTCI (n = 240). Remote estimation of GPP from canopy chlorophyll content × PAR is proved to be relatively successful (R² of 0.47, 0.53, 0.65 and 0.66 for Red edge NDVI, MCARI₇₁₀, CI_{red edge} and MTCI respectively, n = 240). These results open up a new possibility to estimate GPP and should inspire new models for remote sensing of GPP.     

Evapotranspiration and water use efficiency in different-aged Pacific Northwest Douglas-fir stands

We used the eddy-covariance technique to measure evapotranspiration (E) and gross primary production (GPP) in a chronosequence of three coastal Douglas-fir (Pseudotsuga menziesii) stands (7, 19 and 58 years old in 2007, hereafter referred to as HDF00, HDF88 and DF49, respectively) since 1998. Here, we focus on the controls on canopy conductance (g_c), E, GPP and water use efficiency (WUE) and the effect of interannual climate variability at the intermediate-aged stand (DF49) and then analyze the effects of stand age following clearcut harvesting on these characteristics. Daytime dry-foliage Priestley–Taylor α and g_c at DF49 were 0.4–0.8 and 2–6 mm s^?1, respectively, and were linearly correlated (R² = 0.65). Low values of α and g_c at DF49 as well at the other two stands suggested stomatal limitation to transpiration. Monthly E, however, showed strong positive linear correlations to monthly net radiation (R² = 0.94), air temperature (R² = 0.77), and daytime vapour pressure deficit (R² = 0.76). During July–September, monthly E (mm) was linearly correlated to monthly mean soil water content (θ, m³ m^?3) in the 0–60 cm layer (E = 453θ ? 21, R² = 0.69), and GPP was similarly affected. Annual E and GPP of DF49 for the period 1998–2007 varied from 370 to 430 mm and from 1950 to 2390 g C m^?2, respectively. After clearcut harvesting, E dropped to about 70% of that for DF49 while ecosystem evapotranspiration was fully recovered when stand age was ～12 years. This contrasted to GPP, which varied hyperbolically with stand age. Monthly GPP showed a strong positive linear relationship with E irrespective of the stand age. While annual WUE of HDF00 and HDF88 varied with age from 0.5 to 4.1 g C m^?2 kg^?1 and from 2.8 to 4.4 g C m^?2 kg^?1, respectively, it was quite conservative at ～5.3 g C m^?2 kg^?1 for DF49. N-fertilization had little first-year response on E and WUE. This study not only provides important results for a more detailed validation of process-based models but also helps in predicting the influences of climate change and forest management on water vapour and CO₂ fluxes in Douglas-fir forests.     

Microbial utilization and mineralization of [14C]glucose added in six orders of concentration to soil

The substrate availability for microbial biomass (MB) in soil is crucial for microbial biomass activity. Due to the fast microbial decomposition and the permanent production of easily available substrates in the rooted top soil mainly by plants during photosynthesis, easily available substrates make a very important contribution to many soil processes including soil organic matter turnover, microbial growth and maintenance, aggregate stabilization, CO₂ efflux, etc. Naturally occurring concentrations of easily available substances are low, ranging from 0.1 μM in soils free of roots and plant residues to 80 mM in root cells. We investigated the effect of adding ¹⁴C-labelled glucose at concentrations spanning the 6 orders of magnitude naturally occurring concentrations on glucose uptake and mineralization by microbial biomass. A positive correlation between the amount of added glucose and its portion mineralized to CO₂ was observed: After 22 days, from 26% to 44% of the added 0.0009 to 257 μg glucose C g^?1 soil was mineralized. The dependence of glucose mineralization on its amount can be described with two functions. Up to 2.6 μg glucose C g^?1 soil (corresponds to 0.78% of initial microbial biomass C), glucose mineralization increased with the slope of 1.8% more mineralized glucose C per 1 μg C added, accompanied by an increasing incorporation of glucose C into MB. An increased spatial contact between micro-organisms and glucose molecules with increasing concentration may be responsible for this fast increase in mineralization rates (at glucose additions <2.6 μg C g^?1). At glucose additions higher than 2.6 μg C g^?1 soil, however, the increase of the glucose mineralization per 1 μg added glucose was much smaller as at additions below 2.6 μg C g^?1 soil and was accompanied by decreasing portions of glucose ¹⁴C incorporated into microbial biomass. This supports the hypothesis of decreasing efficiency of glucose utilization by MB in response to increased substrate availability in the range 2.6–257 μg C g^?1 (=0.78–78% of microbial biomass C). At low glucose amounts, it was mainly stored in a chloroform-labile microbial pool, but not readily mineralized to CO₂. The addition of 257 μg glucose C g^?1 soil (0.78 μg C glucose μg^?1 C micro-organisms) caused a lag phase in mineralization of 19 h, indicating that glucose mineralization was not limited by the substrate availability but by the amount of MB which is typical for 2nd order kinetics.     

The fate and toxicity of the flavonoids naringenin and formononetin in soil

The flavonoid class of plant secondary metabolites play a multifunctional role in below-ground plant–microbe interactions with their best known function as signals in the nitrogen fixing legume–rhizobia symbiosis. Flavonoids enter rhizosphere soil as a result of root exudation and senescence but little is known about their subsequent fate or impacts on microbial activity. Therefore, the present study examined the sorptive behaviour, biodegradation and impact on dehydrogenase activity (as determined by iodonitrotetrazolium chloride reduction) of the flavonoids naringenin and formononetin in soil. Organic carbon normalised partition coefficients, log K_oc, of 3.12 (formononetin) and 3.19 (naringenin) were estimated from sorption isotherms and, after comparison with literature log K_oc values for compounds whose soil behaviour is better characterised, the test flavonoids were deemed to be moderately sorbed. Naringenin (spiked at 50 μg g^?1) was biodegraded without a detectable lag phase with concentrations reduced to 0.13±0.01 μg g^?1 at the end of the 96 h time course. Biodegradation of formononetin proceeded after a lag phase of ～24 h with concentrations reduced to 4.5±1% of the sterile control after 72 h. Most probable number (MPN) analysis revealed that prior to the addition of flavonoids, the soil contained 5.4×10⁶ MPN g^?1 (naringenin) and 7.9×10⁵ MPN g^?1 (formononetin) catabolic microbes. Formononetin concentration had no significant (p>0.05) effect on soil dehydrogenase activity, whereas naringenin concentration had an overall but non-systematic impact (p=0.045). These results are discussed with reference to likely total and bioavailable concentrations of flavonoids experienced by microbes in the rhizosphere.     

Stimulatory effect of phosphate-solubilizing fungal strains (Aspergillus awamori and Penicillium citrinum) on the yield of chickpea (Cicer arietinum L. cv. GPF2)

The effect of six phosphate-solubilizing fungi (PSF, two strains of Aspergillus awamori, and four of Penicillium citrinum) isolated from rhizosphere of various crops, was observed on the growth and seed production of chickpea plants (Cicer arietinum L. cv. GPF2) in pot experiments. The phosphate (P) solubilizing activity of PSF in liquid varied from 38 to 760 μg ml^?1 for tricalcium phosphate (TCP) and 28–248 μg ml^?1 for mussoorie rock phosphate (MRP). All PSF isolates were biocompatible and produced growth-promoting hormone, Indole acetic acid (IAA), varying in concentration from 2.5 to 9.8 μg ml^?1. Of the various pot experiments carried out in green house, maximum stimulatory effect on chickpea plants growth was observed by inoculation of two A. awamori strains. This treatment resulted in 7–12% increase in shoot height, nearly three-fold increase in seed number and two-fold increase in seeds weight as compared to the control (un-inoculated) plants. Inoculation of four strains of P. citrinum exhibited lesser stimulatory effect. It showed 7% increase in shoot height, two-fold increase in seed number and 87% increase in seeds weight as compared to the control plants. However, a consortium of all the six fungal isolates showed no stimulatory effect on chickpea plants growth.     

Modeling organic carbon dynamics under no-tillage and plowed systems in tropical soils of Brazil using CQESTR

CQESTR simulates the effect of management practices on soil organic carbon (SOC) stocks. The beta version of the model had been calibrated and validated for temperate regions. Our objective was to evaluate the CQESTR model performance for simulating carbon dynamics as affected by tillage practices in two tropical soils (Ultisol and Oxisol) in southeastern and northeastern Brazil. In the southeast (20.75 S 42.81 W), tillage systems consisted of no tillage (NT); reduced tillage (RT) (one disc plow and one harrow leveling [RT1] or one heavy disc harrow and one harrow leveling [RT2]); and conventional tillage (CT) (two heavy disc harrows followed by one disc plow and two harrow levelings). In the northeast (7.55 S 45.23 W), tillage systems consisted of NT, RT (one chisel plow and one harrow leveling), and CT (one disk plow, two heavy disk harrowings, and two harrow levelings). CQESTR underestimated SOC at both sites, especially under NT systems, indicating that adjustments (e.g., the inclusion of clay mineralogy factor) are necessary for more accurate simulation of SOC in the tropics. In spite of this, measured and simulated values of SOC in the 0–20 cm depth were well correlated (southeast, R² = 0.94, p < 0.01; northeast, R² = 0.88, p < 0.05). With respect to initial conditions (native forest), CQESTR estimated a decrease in SOC stocks in plowed and no-tillage systems. In 2006, in the southeast, SOC stocks were 28.8, 23.7, 23.2, and 22.0 Mg ha^?1 under NT, RT2, RT1, and CT, respectively; in the northeast, stocks were 36.0, 33.8, and 32.5 Mg ha^?1 under NT, RT, and CT, respectively. The model estimated carbon emissions varying from 0.36 (NT) to 1.05 Mg ha^?1 year^?1 (CT) in the southeast and from 0.30 (NT) to 0.82 (CT) Mg ha^?1 year^?1 in the northeast. CQESTR prediction of SOC dynamics illustrates acceptable performance for the two tropical soils of Brazil.     

Characteristics of earthworms (Eisenia fetida) in PAHs contaminated soil amended with sewage sludge or vermicompost

Earthworms can be used to remove polycyclic aromatic hydrocarbons (PAHs) from soil, but this might affect their survival and they might accumulate the contaminants. Sterilized and unsterilized soil was contaminated with phenanthrene (Phen), anthracene (Anth) and benzo(a)pyrene (BaP), added with or without Eisenia fetida, sewage sludge or vermicompost. Survival, growth, cocoon formation and concentrations of PAHs in the earthworms were monitored for 70 days. Addition of sewage sludge to sterilized or unsterilized soil maintained the number of earthworms and their survival was 94%. The addition of sludge significantly increased the weight of earthworms 1.3 times compared to those kept in the unamended soil or in soil amended with vermicompost. The weight of earthworms was significantly lower in sterilized than in unsterilized soil. Cocoons were only detected when sewage sludge was added to unsterilized soil. A maximum concentration of 62.3 μg Phen kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost after 7 days and 22.3 μg Phen kg⁻¹ when kept in the unamended unsterilized soil after 14 days. Concentrations of Phen in the earthworms decreased thereafter and ≤2 μg kg⁻¹ after 28 days. A maximum Anth concentration of 82.5 μg kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost and 45.8 μg Anth kg⁻¹ when kept in the unamended unsterilized soil after 14 days. A maximum concentration of 316 μg BaP kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost after 56 days and 311 μg BaP kg⁻¹ when kept in the unsterilized soil amended with vermicompost after 28 days. The amount of BaP in the earthworm was generally largest after 28 days, but after 70 days still 60 μg kg⁻¹ was found in E. fetida when kept in the sterilized soil amended with sewage sludge. It was found that E. fetida survived in PAHs contaminated soil and accumulated only small amounts of the contaminants, but sewage sludge was required as food for its survival and cocoon production.     

Miscanthus sinensis and wild flowers as food resources of Lumbricus terrestris L.

Senescent leaves of Miscanthus sinensis contained 36% soluble polysaccharides, 26% cellulose and had a C/N ratio of 45. In 11 wild flower species contents of soluble polysaccharides (21–30%), cellulose (3–16%) and C/N ratio (13–31) were lower. Decomposing leaves of M. sinensis lost weight at a rate of 0.002 day⁻¹, increased the C/N ratio from 45 to about 100, the bacterial biomass from 0.4 to 1 μg C mg⁻¹ dry weight, and decreased the tensile strength from 35 to 10 N. The withdrawal rate of Lumbricus terrestris with senescent leaves of M. sinensis was 30 mg g⁻¹ week⁻¹; the feeding rate was lower. With most senescent wild flowers withdrawal and feeding rates were higher. During decomposition of M. sinensis withdrawal rates increased to about 90, and feeding rates to about 30 mg g⁻¹ week⁻¹. The rates were not related to soluble polysaccharides, cellulose, acid-insoluble residue, C/N ratio and the presence of trichomes on the leaves. The abundance of L. terrestris decreased in a meadow turned into a field of M. sinensis from 55 to 26 earthworms m⁻² and increased in a rotational maize field turned into wild flower strips from 28 to 46 earthworms m⁻². The species richness of earthworms decreased with M. sinensis from 7.2 to 4.7 and increased with wild flowers from 4.7 to 6.7 species per sampling unit.     

Endogeic and anecic earthworm abundance in six Midwestern cropping systems

Endogeic and juvenile anecic earthworm abundance was measured in soil samples and anecic populations were studied by counting midden numbers at the sites of two long-term cropping systems trials in South-central Wisconsin. The three grain and three forage systems at each site were designed to reflect a range of Midwestern USA production strategies. The primary objectives of this work were to determine if the abundance of endogeic or anecic earthworms varied among cropping systems or crop phases within a cropping system and were there specific management practices that impacted endogeic or anecic earthworm numbers. The earthworms present in the surface soil were: Aporrectodea tuberculata (Eisen), A. caliginosa (Savigny), A. trapezoides (Dugés); and juvenile Lumbricus terrestris (L.). True endogeic abundance was greatest in rotationally grazed pasture [188 m^?2 at Arlington (ARL) and 299 m^?2 at Elkhorn (ELK)], and smallest in conventional continuous corn (27 m^?2 at ARL and 32 m^?2 at ELK). The only type of anecic earthworm found was L. terrestris L. There was an average of 1.2 middens per adult anecic earthworm and the population of anecics was greatest in the no-till cash grain system (28 middens m^?2 at ARL, 18 m^?2 at ELK) and smallest in the conventional continuous corn system (3 middens m^?2 at ARL, 1 m^?2 at ELK). Earthworm numbers in individual crop phases within a cropping system were too variable from year-to-year to recommend using a single phase to characterize a whole cropping system. Indices for five management factors (tillage, manure inputs, solid stand, pesticide use, and crop diversity) were examined, and manure use and tillage were the most important impacting earthworm numbers across the range of cropping systems. Manure use was the most important management factor affecting endogeic earthworm numbers; but no-tillage was the most important for the juvenile and adult anecic groups and had a significantly positive influence on endogeic earthworm counts as well. The pesticides used, which were among the most commonly applied pesticides in the Midwestern USA, and increasing crop diversity did not have a significant effect on either the endogeic or anecic earthworm groups in this study. Consequently, designing cropping systems that reduce tillage and include manure with less regard to omitting pesticides or increasing crop diversity should enhance earthworm populations and probably improve sustainability.     

Tolerance,growth and degradation of phenanthrene and benzo[a]pyrene by Rhizobium tropici CIAT 899 in liquid culture medium

Polycyclic aromatic hydrocarbons (PAH) are ubiquitous pollutants that are toxic and recalcitrant to degradation by bacteria. This research evaluated the toxicity of different concentrations [10, 20, 40, 60, 80 and 100 μg mL⁻¹] of phenanthrene (PHE) or benzo[a]pyrene (BaP) on the growth of Rhizobium tropici CIAT899 under in vitro conditions as well as the potential degradation of PHE and BaP by this bacterium. At 24 h, a 40% decrease in Rhizobium growth was observed when exposed to 40 μg mL⁻¹ of either PHE or BaP. Furthermore, bacterial growth was completely inhibited by PHE or BaP applied in 80 and 100 μg mL⁻¹. After 96 h, the growth of R. tropici at 40 μg PHE mL⁻¹ or 60 μg BaP mL⁻¹ was similar to those treatments without PAH. To evaluate R. tropici degrading capabilities, supernatants of cultures with 40 μg PHE mL⁻¹ or 60 μg BaP mL⁻¹ were analyzed by gas chromatography coupled to mass spectrophotometer (GC–MS). R. tropici was able to degrade either PHE or BaP diminishing its concentration in 20% and 25% during the first 24 h, degradation obtained at 120 h was 50% and 45% for PHE or BaP, respectively. This research shows for the first time that R. tropici CIAT 899 grows in liquid culture medium contaminated with PAH, and moreover is able to growth and to degrade either PHE or BaP.     

Impact of bovine urine deposition on soil microbial activity,biomass, and community structure

Nitrogen (N) from urine excreted by grazing animals can be transformed into N compounds that have detrimental effects on the environment. These include nitrate, which can cause eutrophication of waterways, and nitrous oxide, which is a greenhouse gas. Soil microbes mediate all of these N transformations, but the impact of urine on microbes and how initial soil conditions and urine chemical composition alter their responses to urine are not well understood. This study aimed to determine how soil inorganic N pools, nitrous oxide fluxes, soil microbial activity, biomass, and the community structure of bacteria containing amoA (nitrifiers), nirK, and nirS (denitrifiers) genes responded to the addition of urine over time. Bovine urine containing either a high (15.0 g K⁺ l^?1) or low salt content (10.4 g K⁺ l^?1) was added to soil cores at either low or high moisture content (hereafter termed dry and wet soil respectively; 35% or 70% water-filled pore space after the addition of urine). Changes in soil conditions, inorganic N pools, nitrous oxide fluxes, and the soil microbial community were then measured 1, 3, 8, 15, 29 and 44 days after urine addition. Urine addition increased soil ammonium concentrations by up to 2 mg g d.w.^?1, soil pH by up to 2.7 units, and electrical conductivity (EC) by 1.0 and 1.6 dS m^?1 in the low and high salt urine treatments respectively. In response, nitrate accumulation and nitrous oxide fluxes were lower in dry compared to wet urine-amended soils and slightly lower in high compared to low salt urine-amended soils. Nitrite concentrations were elevated (>3 μg g d.w.^?1) for at least 15 days after urine addition in wet urine-amended soils, but were only this high in the dry urine-amended soils for 1 day after the addition of urine. Microbial biomass was reduced by up to half in the wet urine-amended soils, but was largely unaffected in the dry urine-amended soils. Urine addition affected the community structure of ammonia-oxidising and nitrite-reducing bacteria; this response was also stronger and more persistent in wet than in dry urine-amended soils. Overall, the changes in soil conditions caused by the addition of urine interacted to influence microbial responses, indicating that the effect of urine on soil microbes is likely to be context-dependent.     

Enzyme activities and diuron persistence in soil amended with vermicompost derived from spent grape marc and treated with urea

Mineral fertilizers, organic amendments, and pesticides are inputs commonly used in conventional farming practices. The aim of this study was to evaluate the effects of single or combined applications of spent grape marc-vermicompost, urea, and/or diuron on soil-enzyme activities and the persistence of this herbicide in soils with low organic carbon content. The application of vermicompost enhanced dehydrogenase (DHase) enzyme activity over time but altered soil urease activity to a very limited extent. The reduction in diuron concentrations and the increase in DHase activity indicated that the soil microorganisms were capable of degrading the ureic herbicide. Treatment with vermicompost and diuron had a stimulatory effect on soil microbial activity. On the whole, the application of diuron and urea to the vermicompost-amended soil raised DHase and urease activity to maximum levels (>3 μg INTF g^?1 h^?1 and >47 μg NH₄⁺ g^?1 h^?1, respectively). The application of urea to the unamended and vermicompost-amended soil decreased diuron persistence from 18.8 and 33 d to 12.5 and 15 d, respectively. Our findings show that although vermicompost additions reduce diuron availability, this boosts diuron degradation when combined with urea. These additions, under different soil management conditions, minimize the bioavailability and persistence of diuron and consequently the risk of leaching and seepage into aquifers. Compared with untreated soils, these types of treated soils could also improve agricultural sustainability and the quality of the environment.     

Multiple measurements constrain estimates of net carbon exchange by a Eucalyptus forest

Net ecosystem exchange of carbon (F_NEE) was estimated for a temperate broadleaf, evergreen eucalypt forest ecosystem at Tumbarumba in south-eastern Australia to investigate the processes controlling forest carbon sinks and their response to climate. Measurements at a range of temporal and spatial scales were used to make three different estimates of F_NEE based on: (1) the difference between fluxes of carbon input by photosynthesis and output by autotrophic plus heterotrophic respiration, (2) changes over time in the carbon pools in the above- and below-ground biomass, soil and litter, and (3) micrometeorological flux measurements that provide a continuous estimate of the net exchange. A rigorous comparison of aggregated component fluxes and the net eddy fluxes within a flux tower source area was achieved based on an inventory of the site and a detailed sampling strategy. Measurements replicated in space and time provided mean values, confidence limits and patterns of variation of carbon pools and fluxes that allowed comparisons within known limits of uncertainty. As a result of comparisons between nighttime eddy flux and chamber measurements of respiration, a revised micrometeorological method was developed for estimating nighttime carbon flux using flux tower measurements. Uncertainty in the final estimate of F_NEE was reduced through mutual constraints of each of these measurement approaches. F_NEE for the period October 2001–September 2002, with average rainfall, was an uptake of 6.7 (5.1–8.3) tC ha^?1 yr^?1 estimated from component fluxes, and 5.4 (3.0–7.5) tC ha^?1 yr^?1 estimated from the revised eddy flux method. Biomass increment was 4.5 (3.7–5.4) tC ha^?1 yr^?1 and the remaining 0.9–2.2 tC ha^?1 yr^?1 could represent a carbon sink in the soil and litter pools or lie within the confidence limits of the measured fluxes. F_NEE was reduced to ?0.1 to 2.4 tC ha^?1 yr^?1 during a period of drought and insect disturbance in October 2002–September 2003, with biomass increment being the main component reduced. The forest is a large carbon sink compared with other forest ecosystems, but this is subject to high-annual variability in response to climate variability and disturbance.     

Field margins and management affect settlement and spread of an introduced dew-worm (Lumbricus terrestris L.) population

To study the feasibility of earthworm introduction for increasing the macroporosity and permeability of arable heavy clay, deep-burrowing earthworms (Lumbricus terrestris L.) were inoculated into a tile drained experimental field in Jokioinen, S-W Finland in autumn 1996. Inoculation with the Earthworm Inoculation Unit technique was at the up-slope end of the field, in the field margins under permanent grass, and inside the four 0.46 ha plots of the field. The experiment was monitored on three occasions. In 1998 the L. terrestris population had persisted in low numbers only in field and plot margins. By 2003, when the field had been under set-aside grass for three years, density had grown in the margins and L. terrestris were also found inside the field at a very low density. The third monitoring was in autumn 2009, after a further four years as set-aside and a subsequent division of the field into no-till and ploughing management, and looked at the effects of management (margins, no-till, ploughing), distance from the inoculation and sub-drainage on L. terrestris abundance. The abundance displayed a clear gradient over the field, declining from 14 ind. and 18 g m^?2 at 5–9 m from inoculation, to 1 ind. and 2 g m^?2 at 56–60 m distance. Margins had the highest abundances (16 ind. and 32 g m^?2), followed by no-till (4 ind. and 4 g m^?2) and ploughing (1 ind. and 1 g m^?2). Abundances were significantly higher above the tiles than between them (P < 0.05). The results demonstrate the importance of no-till and sub-drain line habitats as settlement supports for the inoculated population. Field margins proved to be decisive for inoculation success, by providing bridgeheads for population establishment and later by acting as source areas for the colonisation of the field. This finding highlights the general importance of field margins in the dispersal ecology of earthworms in arable landscapes.     

Rapid estimation of microbial biomass in grassland soils by ultra-violet absorbance

A reliable and simple technique for estimating soil microbial biomass (SMB) is essential if the role of microbes in many soil processes is to be quantified. Conventional techniques are notoriously time-consuming and unreproducible. A technique was investigated that uses the UV absorbance at 280 nm of 0.5 M K₂SO₄ extracts of fumigated and unfumigated soils to estimate the concentrations of carbon, nitrogen and phosphorus in the SMB. The procedure is based on the fact that compounds released after chloroform fumigation from lysed microbial cells absorb in the near UV region. Using 29 UK permanent grassland soils, with a wide range of organic matter (2.9–8.0%) and clay contents (22–68%), it was demonstrated that the increase in UV absorbance at 280 nm after soil fumigation was strongly correlated with the SMB C (r=0.92), SMB N (r=0.90) and SMB P (r=0.89), as determined by conventional methods. The soils contained a wide range of SMB C (412–3412 μg g⁻¹ dry soil), N (57–346 μg g⁻¹ dry soil) and P (31–239 μg g⁻¹ dry soil) concentrations. It was thus confirmed that the UV absorbance technique described was a rapid, simple, precise and relatively inexpensive method of estimating soil microbial biomass.     

Detection and quantification of the nematophagous fungus Hirsutella minnesotensis in soil with real-time PCR

A real-time PCR assay was developed to quantify in soil the fungus Hirsutella minnesotensis, an important parasite of secondary-stage juvenile (J2) of the soybean cyst nematode. A primer pair 5′-GGGAGGCCCGGTGGA-3′ and 5′-TGATCCGAGGTCAACTTCTGAA-3′ and a TaqMan probe 5′-CGTCCGCCGTAAAACGCCCAAC-3′ were designed based on the sequence of the ITS region of the rRNA gene. The primers were highly species-specific. The PCR reaction system was very sensitive and able to detect as few as 4 conidia g^?1 soil. Regression analysis showed similar slopes and efficiency on DNA from pure culture (y = ?3.587x + 41.017, R² = 0.9971, E = 0.9055) and from Log conidia g^?1 soil (y = ?3.855x + 37.669, R² = 0.9139, E = 0.8172), indicating that the real-time PCR protocol can reliably quantify H. minnesotensis in the soil. The real-time PCR assay was applied to 20 soil samples from soybean fields, and compared with a parasitism assay. The real-time PCR assay detected H. minnesotensis in six of the soils, whereas the parasitism assay detected H. minnesotensis in the same six soils and three additional soils. The real-time PCR assay was weakly correlated (R² = 0.49) with the percentage of parasitized J2 in the six soils, indicating that different types of soil may interfere the efficiency of the real-time PCR assay, possibly due to the effect of soil types on efficacy of DNA extraction. The parasitism assay appeared to be more sensitive than real-time PCR in detecting presence of H. minnesotensis, but real-time PCR was much faster and less costly and provided a direct assessment of fungal biomass. Using the two assays in combination can obtain more complete information about the fungus in soil than either assay alone. Hirsutella parasitism was widespread and detected in 13 of the 20 field soils, indicating that these fungi may contribute to suppressiveness of soybean cyst nematode in nature and likely have high biological control potential for the nematode.     

Comparison of the eddy covariance and automated closed chamber methods for evaluating nocturnal CO2 exchange in a Japanese cypress forest

Underestimation of nocturnal CO₂ respiration using the eddy covariance method under calm conditions remains an unsolved problem at many flux observation sites in forests. To evaluate nocturnal CO₂ exchange in a Japanese cypress forest, we observed CO₂ flux above the canopy (F_c), changes in CO₂ storage in the canopy (S_t) and soil, and trunk and foliar respiration for 2 years (2003–2004). We scaled these chamber data to the soil, trunk, and foliar respiration per unit of ground area (F_s, F_t, F_f, respectively) and used the relationships of F_s, F_t, and F_f with air or soil temperature for comparison with canopy-scale CO₂ exchange measurements (=F_c + S_t). The annual average F_s, F_t, and F_f were 714 g C m⁻² year⁻¹, 170 g C m⁻² year⁻¹, and 575 g C m⁻² year⁻¹, respectively. At small friction velocity (u_*), nocturnal F_c + S_t was smaller than F_s + F_t + F_f estimated using the chamber method, whereas the two values were almost the same at large u_*. We replaced F_c + S_t measured during calm nocturnal periods with a value simulated using a temperature response function derived during well-mixed nocturnal periods. With this correction, the estimated net ecosystem exchange (NEE) from F_c + S_t data ranged from −713 g C m⁻² year⁻¹ to −412 g C m⁻² year⁻¹ in 2003 and from −883 g C m⁻² year⁻¹ to −603 g C m⁻² year⁻¹ in 2004, depending on the u_* threshold. When we replaced all nocturnal F_c + S_t data with F_s + F_t + F_f estimated using the chamber method, NEE was −506 g C m⁻² year⁻¹ and −682 g C m⁻² year⁻¹ for 2003 and 2004, respectively.