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.     

Toxicity of three pesticides commonly used in Brazil to Pontoscolex corethrurus (Müller, 1857) and Eisenia andrei (Bouché, 1972)

The indiscriminate and excessive use of pesticides poses serious risks to humans and the environment, including soil biota. Ecotoxicological tests are useful to indicate the extent to which these chemicals are harmful and how and where their effects occur. Some of these tests were standardized by ISO (International Organization for Standartization) using the earthworm species Eisenia fetida and Eisenia andrei, both native to temperate climates. However, these species may be of lower relevance for soil ecotoxicological studies since they live in the litter and feed on fresh organic matter. The species Pontoscolex corethrurus, native to tropical regions, may be an alternative for more relevant ecotoxicological tests as it is an endogeic geophagous species. However, little is known of its sensitivity to pesticides. Therefore, avoidance and mortality tests were performed using E. andrei and P. corethrurus and three pesticides commonly used in Brazilian agriculture: carbendazim, carbofuran and glyphosate. The tests were conducted in tropical artificial soil (TAS). For carbendazim, the median avoidance concentration (AC₅₀) was 76.1 and 65.8 mg a.i. kg⁻¹ and the median lethal concentration (LC₅₀) 19.7 and 15.3 mg a.i. kg⁻¹ for E. andrei and P. corethrurus, respectively. For carbofuran, the AC₅₀ was 9.7 and 7.3 mg a.i. kg⁻¹ and LC₅₀ 13.5 and 9.3 mg a.i. kg⁻¹ for E. andrei and P. corethrurus, respectively. Concentrations applied in the field of these two pesticides have toxic effects on both species. Glyphosate showed no toxic effects for either species even at the highest concentration tested (47 mg a.i. kg⁻¹), although they displayed avoidance behavior at this concentration. The sensitivity of P. corethrurus appears to be similar to the standard species for the pesticides evaluated reinforcing the notion that E. andrei is a good test species. Nevertheless, further studies should be undertaken using other contaminants to confirm the similar sensitivity of both species and the relevance of E. andrei in ecotoxicological tests.     

Application of biostimulants in benzo(a)pyrene polluted soils: Short-time effects on soil biochemical properties

The bioremediation effects of three biostimulants (BS): WCDS, wheat condensed distillers soluble; PAHE, hydrolyzed poultry feathers; and RB, rice bran extract in a soil polluted with two rates of benzo(a)pyrene (BaP) (50 or 100 mg kg⁻¹ soil, respectively) over 90 days were studied. Their effects on the soil biochemical properties (ATP and urease and phosphatase activities) and ergosterol were determined. Also, extractable BaP in soils was determined during the incubation period. An non-polluted and non-organic-amended soil was used as control. The results indicated that at the end of the incubation period and compared with the control soil, the ATP, ergosterol, urease and phosphatase activities decreased 29.4%, 24.8%, 44.7% and 42.9%, respectively in the non-organic amended soil polluted with polluted 100 mg BaP kg⁻¹ soil. The application of biostimulants to unpolluted soil increased the biochemical parameters. However, this stimulation was higher in the soil amended with PAHE, followed by RB and WCDS. The application of BaP in organic-amended soils decreased the biochemical properties. However, this decrease was lower than for the non-amended BaP polluted soil. Possibly the low molecular weight protein content easily assimilated by soil microorganisms is responsible for less inhibition of these soil biochemical parameters.     

Effects of azoxystrobin,chlorothalonil, and ethoprophos on the reproduction of three terrestrial invertebrates using a natural Mediterranean soil

The potential terrestrial toxicity of three pesticides, azoxystrobin, chlorothalonil, and ethoprophos was evaluated using reproduction ecotoxicological tests with different non-target species: the collembolan Folsomia candida, the earthworm Eisenia andrei, and the enchytraeid Enchytraeus crypticus. All reproduction tests were performed with natural soil from a Mediterranean agricultural area (with no pesticide residues) in order to improve the relevance of laboratory data to field conditions. Controls were performed with natural and standard artificial soil (OECD 10% OM). The fungicide azoxystrobin showed the highest toxicity to earthworms (EC₅₀ = 42.0 mg a.i. kg⁻¹ dw soil). Collembolans were the most sensitive taxa in terms of sublethal effects of chlorothalonil with an EC₅₀ of 31.1 mg a.i. kg⁻¹ dw soil followed by the earthworms with an EC₅₀ of 40.9 mg a.i. kg⁻¹ dw soil. The insecticide ethoprophos was the most toxic to collembolans affecting their reproduction with an EC₅₀ of 0.027 mg a.i. kg⁻¹ dw soil. Enchytraeids were generally the least sensitive of the three species tested for long-term effects. Earthworms were not always the most sensitive species, emphasizing the need to increase the number of mandatory assays with key non-target organisms in the environmental risk assessment of pesticides.     

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.     

Enhancement of arbuscular mycorrhizal fungus (Glomus versiforme) on the growth and Cd uptake by Cd-hyperaccumulator Solanum nigrum

Arbuscular mycorrhizal fungus (AMF) can enhance plant growth and resistance to toxicity produced by heavy metals (HMs), affect the bioavailability of HMs in soil and the uptake of HMs by plants, and thus has been emerged as the most prominent symbiotic fungus for contribution to phytoremediation. A greenhouse pot experiment was conducted to assess the effect of Glomus versiforme BGC GD01C (Gv) on the growth and Cd accumulation of Cd-hyperaccumulator Solanum nigrum in different Cd-added soils (0, 25, 50, 100 mg Cd kg⁻¹ soil). Mycorrhizal colonization rates were generally high (from 71% to 82%) in Gv-inoculated treatments at all Cd levels. Gv colonization enhanced soil acid phosphatase activity, and hence elevated P acquisition and growth of S. nigrum at all Cd levels. Moreover, the presence of Gv significantly increased DTPA-extractable (phytoavailable) Cd concentrations in 25 and 50 mg Cd kg⁻¹ soils, but did not affect phytoavailable Cd in 100 mg Cd kg⁻¹ soil. Similarly, inoculation with Gv significantly increased Cd concentrations of S. nigrum in 25 and 50 mg Cd kg⁻¹ soils, but decreased Cd concentrations of the plants in 100 mg Cd kg⁻¹ soil. Overall, inoculation with Gv greatly improved the total Cd uptakes in all plant tissues at all Cd levels. The present results indicated that S. nigrum associated with Gv effectively improved the Cd uptake by plant and would be a new strategy in microbe-assisted phytoremediation for Cd-contaminated soils.     

Molecular and physiological bacterial diversity of a semi-arid soil contaminated with different levels of formulated atrazine

A semi-arid soil treated with different concentrations of formulated atrazine in a laboratory experiment was studied over 45 days, by different biological and molecular parameters (bacterial enumeration (cfu), community level physiological profiles (CLPPs) measured by Biolog^® and denaturing gradient gel electrophoresis (DGGE)), to study the bacterial community diversity.Formulated atrazine was almost totally degraded at different concentrations after this incubation time. The number of colony forming units (cfu) for soils with 100 and 1000 mg kg⁻¹ atrazine was significantly (p ≤ 0.05) higher than for the control, 1 and 10 mg kg⁻¹ treatments. DGGE banding patterns showed that regardless of time elapsed, concentrations of 10, 100 and 1000 mg kg⁻¹ atrazine in soil, affected the bacterial community compared to control and 1 mg kg⁻¹.The Shannon diversity index (H′) based on CLPP data showed a significant (p ≤ 0.05) decrease at atrazine concentrations of 100 and 1000 mg kg⁻¹. The Shannon diversity indices for different guilds of source carbon and the parameters K and r (based on the kinetics of colour formation rather than on the degree of colour development) were related to guilds of carbon substrates and atrazine concentration at a sampling time. The parameter K was very sensitive to atrazine effects on microbial communities.These biological and molecular parameters can be used to monitor changes in soils treated with atrazine at different concentrations, even when the pesticide is degraded.     

Soil available phosphorus status determines indigenous mycorrhizal colonization of field and glasshouse-grown spring wheat from Argentina

Wheat production (Triticum aestivum L.) has increased across the world during last century with the intensification of agriculture. Phosphorus (P) fertilization is a common practice to improve wheat growth in Argentina. We investigate whether indigenous arbuscular mycorrhizal colonization (AMC) of hard red spring wheat is controlled by shoot P content (SPc) or by available soil P in an agricultural soil from the southeastern Argentine Pampas. In the field, AMC was monitored four times during two growing seasons of a conventional wheat crop. Treatments were: without P supply, annual supply of 11 and 22 kg P ha⁻¹ during the last 5 years, and 164 kg P ha⁻¹ applied once 5 years before the experiment. In the glasshouse, AMC was assessed three times in wheat growing in pots filled with the soil from unfertilized plots; treatments were: P (0 and 20 mg P pot⁻¹), and nitrogen (N) fertilization (0 and 150 mg N pot⁻¹). A range of soil P between 6 and 60 mg P kg⁻¹ was obtained and the AMC ranged from 1% to 67% of root length colonized under both field and glasshouse conditions. P supplied annually increased growth and SPc but decreased AMC. N fertilization did not affect growth or AMC. Variations in SPc did not account for AMC. Variability in AMC was best accounted for local current soil available P content (r² = 0.59). A linear-plateau relationship between soil P and indigenous AMC was established in wheat plants growing under contrasting environmental and experimental (field and glasshouse) conditions. Indigenous AMC was depressed by available soil P in the range 0–27 mg P kg⁻¹ (a decrease of 2.8% mg P⁻¹ kg⁻¹). Above 27 mg P kg soil⁻¹, AMC was stabilized at about 10%. Grain yield increased with fertilization and the highest relative shoot dry matter in field was obtained at 15.5 mg P kg soil⁻¹. The soil P range that ensures high wheat production without deterring indigenous AMC is discussed.     

Impact of high cadmium and nickel soil concentration on selected physiological parameters of Arundo donax L.

The purpose of this study was to investigate the effects of high cadmium and nickel soil concentrations on selected physiological parameters of Arundo donax L. A 2-year pot experiment was held in the field and the pots were irrigated with aqueous solutions of Cd and Ni in concentrations of 5, 50 and 100 ppm, against the control (tap water). At the end of the cultivation periods the pots soil was divided into three equal zones and total and NH₄OAc extractable Cd and Ni concentrations were determined. The top zone exhibited the highest metal content. Cadmium and nickel total concentrations at the end of the experiment were up to 973.8 mg kg⁻¹ and 2543.3 mg kg⁻¹ respectively, while NH₄OAc extractable Cd was up to 291.7 mg kg⁻¹ and Ni up to 510.3 mg kg⁻¹. Stomatal conductance ranged between 0.3 and 0.8 mol CO₂ m⁻² s⁻¹, intercellular CO₂ concentration ranged between 212.9 and 243.0 ppm CO₂, stomatal resistance between 0.6 and 1.3 s cm⁻¹, chlorophyll content (SPAD values) between 46.3 and 57.0 and chlorophyll fluorescence (F_v/F_m) ranged between 0.8 and 0.9. All studied physiological parameters did not show statistically significant differences among control and heavy metal treated plants for both years; therefore, high soil cadmium and nickel concentration did not inhibit stomatal opening and did not affect the function of the photosynthetic machine of A. donax plants.     

Mercury affects the distribution of culturable species of Pseudomonas in soil

Pseudomonas bacteria isolated during 52 days on Gould's S1 agar from soil spiked with 0, 3.5 and 15 mg Hg(II) kg soil⁻¹ were characterised to reveal whether mercury affected them differently. Isolates from the treatments with 0 and 15 mg Hg kg⁻¹ were characterised using FT-IR characterisation and subsequent 16S rDNA partial sequencing of representative isolates. To verify the selectivity of Gould's S1 agar and the FT-IR characterisation, all 450 isolates were subjected to the following tests: Gram-determination, catalase and oxidase activity, pigment production on PDA and growth at different temperatures. Furthermore, the isolates were tested for their ability to grow on agar amended with 10 mg Hg kg⁻¹ as an indication of mercury resistance. We found that up to 80% of the isolates in soil amended with 15 mg Hg kg⁻¹ were mercury-resistant, whereas only up to 20% were resistant in the treatments with 0 and 3.5 mg Hg kg⁻¹. We found two groups of Pseudomonas, which probably represent non-described species since they did not group closely with any known species of Pseudomonas in the dendrogram. Hg-enhanced isolates were closely related to P. frederiksbergensis. Furthermore, Hg resistance was almost exclusively restricted to P. frederiksbergensis and P. migulae groups. We conclude that Hg caused a shift in the dominating species of culturable Pseudomonas.     

Microbial biomass in a semi arid soil of the central highlands of Mexico cultivated with maize or under natural vegetation

Microbial biomass (MB) is the key factor in nutrient dynamics in soil, but no information exists how clearing of vegetation to cultivate maize in the central highlands of Mexico might affect it. Soil MB was measured with the chloroform fumigation incubation (CFI) and fumigation extraction (CFE) techniques and the substrate-induced respiration (SIR) method in soil sampled under or outside the canopy of mesquite (Prosopis laevigata) and huisache (Acacia tortuoso), N₂ fixing shrubs, and from fields cultivated with maize. Microbial biomass C as measured with the CFI technique ranged from 122 mg C kg⁻¹ in agricultural soil to 373 mg C kg⁻¹ in soil sampled under mesquite shrubs. Microbial biomass N as measured with the CFI technique ranged from 11 mg N kg⁻¹ in agricultural soil to 116 mg N kg⁻¹ in soil sampled under mesquite shrub. The ratio of microbial biomass C as measured with CFI related to the ninhydrin-positive compounds (NPC) was 12.23 after 1 day and 8.43 after 10 days while the relationship with extractable C was 3.15 and 2.96, respectively. The metabolic quotient (qCO₂) decreased in the order OUTSIDE > MESQUITE > HUIZACHE > AGRICULTURE, and the microbial biomass:soil organic C ratio decreased in the order MESQUITE > HUIZACHE > OUTSIDE > AGRICULTURE using SIR to determine the microbial biomass. It was found that converting soil under natural vegetation to arable soil was not only detrimental for soil quality, but might be unsustainable as organic matter input is limited.     

Earthworm-induced N2O emissions in a sandy soil with surface-applied crop residues

Earlier research with endogeic and epigeic earthworm species in loamy arable soil has shown that both earthworm groups can increase nitrous oxide (N₂O) emissions, provided that crop residue placement matches the feeding strategy of the earthworm ecological group(s). However, it is not yet clear whether these effects also occur in sandy soils which typically contain less soil organic matter and have low soil aggregation levels. Here, we aimed to quantify N₂O emissions as affected by endogeic and/or epigeic earthworm species, and to relate changes in N₂O emissions to earthworm-induced changes in soil properties in a sandy soil. A 90 day mesocosm study was conducted with sandy soil and ¹⁵N-labeled radish (Raphanus sativus cv. Adagio L.) residue applied on top. Treatments included: (i) no earthworm addition, (ii) addition of the endogeic species Aporrectodea caliginosa (Savigny), (iii) addition of the epigeic species Lumbricus rubellus (Hoffmeister), and (iv) both species combined. An additional treatment was included without earthworms and with residue manually incorporated into the soil. L. rubellus significantly increased cumulative N₂O emissions from 228 to 859 μg N₂O–N kg^?1 (F_1,12 = 83.12, P < 0.001), whereas A. caliginosa did not affect N₂O emissions. In contrast to earlier studies in loamy soil, no positive interaction between both species with regard to N₂O emissions was found. This was probably related to high competition for organic resources in the relatively poor soil and a low potential for stable soil aggregate formation (and associated anaerobic microsites) by endogeic worms in sandy soil. ¹⁵N isotope analysis revealed that the activity of L. rubellus significantly increased (F_1,12 = 6.20, P = 0.028) the recovery of ¹⁵N in the 250–8000 μm size fraction, indicating incorporation of crop residues into the mineral soil. When residues were manually incorporated, N₂O emissions were significantly (P < 0.008) lower (509 μg N₂O–N kg^?1) than when incorporated by L. rubellus. The high N₂O emissions in the presence of L. rubellus, when compared to manual mixing, suggest a stimulation of microbial activity and/or changes in the microbial community composition. Insights on the earthworm effects on N₂O emission from such soils are discussed.     

Allyl isothiocyanate: an alternative chemical expellant for sampling earthworms

Allyl isothiocyanate (AITC), a natural breakdown product of glucosinolates in many Cruciferae and a component imparting the sharp taste to mustard, was tested for its effectiveness as a chemical expellant for sampling earthworms. Testing was performed in an arable field with earthworm populations dominated by Lumbricus terrestris Linnaeus, 1758 and Aporrectodea tuberculata (Eisen, 1874). The optimal concentration was found by comparing concentrations ranging from 5 to 250 mg l⁻¹ in water. Total biomass and numbers of earthworms collected increased hyperbolically with increasing AITC concentration, with the highest biomass and numbers collected using 100 mg l⁻¹ AITC. Biomass and numbers of earthworms collected using 250 mg l⁻¹ AITC, but not 150 or 200 mg l⁻¹ AITC, were significantly less than with 100 mg l⁻¹ AITC. Less earthworm biomass was collected by hand sorting than with chemical expulsion using 100 mg l⁻¹ AITC, but the number of earthworms collected by the two methods were not different. A comparison of hand sorting and 100 mg l⁻¹ AITC expulsion using analysis of similarities (ANOSIM) showed that the two methods produced samples differing in the distribution of both numbers and biomass of species-by-size classes. Hand sorting collected more of the smallest size class of L. terrestris and the largest size classes of A. tuberculata than AITC expulsion, whereas AITC expulsion recovered more of the largest size classes of L. terrestris than hand sorting. When 100 mg l⁻¹ AITC expulsion was compared with chemical expulsion using 200 mg l⁻¹ formalin, no differences were found in the total number, total biomass or in the species-by-size class distribution of the earthworms collected. This suggests that the AITC method may be a favorable alternative to formalin expulsion for sampling earthworms. Further studies under other environmental conditions and with other species of earthworms are warranted to establish its general utility.     

Arbuscular mycorrhizal abundance in contaminated soils around a zinc and lead deposit

In order to study the variations in spore abundance and root colonization parameters of arbuscular mycorrhizal (AM) fungi in a naturally heavy metals polluted site and their relationships with soil properties, 35 plots in the Anguran Zn and Pb mining region were selected along a transect from the mine to 4500 m away. Within each plot, a composite sample of root and rhizospheric soil from a dominant indigenous plant was collected. The soil samples were analyzed for their physico-chemical characteristics. Spores were extracted, counted and identified at genus level. The roots were examined for colonization, arbuscular abundance, mycorrhizal frequency and intensity. Along the transect, the total and available (DTPA-extractable) concentration of Zn decreased from 6472 to 45 mg kg⁻¹ and 75 to 5 mg kg⁻¹, respectively. For Pb the values varied from 5203 to 0 mg kg⁻¹ and 32 to 0 mg kg⁻¹, respectively. In parallel, root colonization rate in the dominant native plants (except Alyssum sp.) varied from 35% to 85% and the spore numbers from 80 to 1306 per 200 g dry soil along the transect. Spores of Glomus were abundantly found in all plots as dominant, while Acaulospora spores were observed only in some moderately polluted and in control plots. AM fungal propagules never disappeared completely even in soils with the highest rates of both heavy metals. Spore numbers were more affected by Zn and Pb concentrations than root colonization. The variations of AM fungi propagules were better related to available than to total concentration of both metals. Spore numbers were positively correlated with mycorrhizal colonization parameters, particularly with arbuscular abundance.     

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.     

Effects of cultivation on soil microbiological properties in a freshwater marsh soil in Northeast China

The Sanjiang Plain has become an intensive area of land use/cover change in China. However, little is known about the effect of cultivation on soil microbiological properties in this freshwater marsh ecosystem. Our objective was to evaluate the effect of cultivation on mineralizable, microbial biomass, and total C in the Sanjiang Plain of Northeast China. Soil microbial biomass C (MBC) was 4346 ± 309 mg kg⁻¹ in undisturbed marsh and 229 mg kg⁻¹ in soil cultivated for 15 years. Undisturbed marsh soil had the highest microbial quotient (3.64%), which declined with increasing cultivation time (R² = 0.97, p < 0.01). Metabolic quotient increased with increasing cultivation time. Soil C mineralization in undisturbed marsh was 3.5 times that in soil cultivated for 1 year, and was 12 times that in soil cultivated for 15 years. Cultivation strongly affected measured soil microbiological properties.     

A genotype dependent-response to cadmium contamination in soil is displayed by Pinus pinaster in symbiosis with different mycorrhizal fungi

Soil contamination with Cd is of primary concern and beneficial soil restoration strategies urge. The aim of this work is to evaluate the response of two different genotypes of Pinus pinaster (wild and selected) to Cd contamination and to assess how inoculation with ectomycorrhizal fungi, Suillus bovinus and Rhizopogon roseolus, influenced each genotype. Seedlings were exposed to soil contaminated at 15 and 30 mg Cd kg⁻¹. Plant growth, mycorrhizal traits and Cd accumulation in different tissues of the plant were determined at harvest. The fungal community was assessed by denaturing gradient gel electrophoresis. At 15 mg Cd kg⁻¹ S. bovinus increased aboveground development in both genotypes. At 30 mg Cd kg⁻¹ non-inoculated wild genotype accumulated more Cd in the shoots (1.7-fold) than the selected genotype; inoculation with R. roseolus decreased Cd concentration in the roots of the selected genotype whereas the opposite occurred in the wild genotype. Cd concentration in the root system was the parameter most influenced by the interaction between the three studied variables. The fungal community established was affected by the Cd concentration in the soil. Results show that different genotypes of P. pinaster react differently to Cd exposure depending on the mycorrhizal association. The importance of considering the combination between plant genotype and its symbiotic partners when aiming at the forestation of degraded land is highlighted.     

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.     

Earthworm colonisation of abandoned arable soil polluted by copper

The distribution, density and biomass of earthworms were investigated at the copper polluted site, Hygum (Denmark). In 1994, shortly after farming of the area was abandoned, only four earthworm species were present and their distribution was restricted to areas where copper concentration did not exceed 200 mg kg^?1 dry soil. Sixteen years later (in 2010), without any agricultural activity, ten species of earthworms were found, in particular, epigeic species were present where soil copper concentrations reached >1000 mg kg^?1 dry soil.     

Effect of tillage on earthworms over short- and medium-term in conventional and organic farming

Earthworms play an important role in many soil functions and are affected by soil tillage in agricultural soils. However, effects of tillage on earthworms are often studied without considering species and their interactions with soil properties. Furthermore, many field studies are based on one-time samplings that do not allow for characterisation of temporal variation. The current study monitored the short (up to 53 days) and medium term (up to 4 years) effects of soil tillage on earthworms in conventional and organic farming. Earthworm abundances decreased one and three weeks after mouldboard ploughing in both conventional and organic farming, suggesting direct and indirect mechanisms. However, the medium-term study revealed that earthworm populations in mouldboard ploughing systems recovered by spring. The endogeic species Aporrectodea caliginosa strongly dominated the earthworm community (76%), whereas anecic species remained <1% of all earthworms in all tillage and farming systems over the entire study. In conventional farming, mean total earthworm abundance was not significantly different in reduced tillage (153 m⁻²) than mouldboard ploughing (MP; 130 m⁻²). However, reduced tillage in conventional farming significantly increased the epigeic species Lumbricus rubellus from 0.1 m⁻² in mouldboard ploughing to 9 m⁻² averaged over 4 years. Contrastingly, in organic farming mean total earthworm abundance was 45% lower in reduced tillage (297 m⁻²) than MP (430 m⁻²), across all sampling dates over the medium-term study (significant at 3 of 6 sampling dates). Reduced tillage in organic farming decreased A. caliginosa from 304 m⁻² in mouldboard ploughing to 169 m⁻² averaged over 4 years (significant at all sampling dates). Multivariate analysis revealed clear separation between farming and tillage systems. Earthworm species abundances, soil moisture, and soil organic matter were positively correlated, whereas earthworm abundances and penetration resistance where negatively correlated. Variability demonstrated between sampling dates highlights the importance of multiple samplings in time to ascertain management effects on earthworms. Findings indicate that a reduction in tillage intensity in conventional farming affects earthworms differently than in organic farming. Differing earthworm species or ecological group response to interactions between soil tillage, crop, and organic matter management in conventional and organic farming has implications for management to maximise soil ecosystem functions.