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.     

Removal of benzo (a) pyrene from soil using an endogeic earthworm Pontoscolex corethrurus ()

The endogeic earthworm Pontoscolex corethrurus (Müller, 1857) was the most abundant species (75%) in soil contaminated with hydrocarbons, mostly benzo(a)pyrene (BaP), in the state of Tabasco (Mexico). The earthworm P. corethrurus was tested for its capacity to remove 100 mg BaP kg⁻¹ from an Anthrosol soil (sterilized or not) and amended with legume Mucuna pruriens (L.) DC. var. utilis (Wall. ex Wight) Baker ex Burck (3%) or the grass Brachiaria humidicola (L.) DC (3%) (recently renamed as Urochloa humidicola (Rendle) Morrone & Zuloaga) in an aerobic incubation experiment. P. corethrurus removed 26.6 mg BaP kg⁻¹ from the sterilized soil and application of B. humidicola as feed increased this to 35.7 mg BaP kg⁻¹ and M. pruriens to 34.2 mg BaP kg⁻¹ after 112 days. The autochthonous microorganisms removed 9.1 mg BaP kg⁻¹ from the unsterilized soil and application of B. humidicola increased this to 18.0 mg BaP kg⁻¹ and M. pruriens to 11.2 mg BaP kg⁻¹. Adding P. corethrurus to the unsterilized soil accelerated the removal of BaP and 36.1 mg kg⁻¹ was dissipated from soil. It was found that the autochthonous microorganisms removed BaP from soil, but addition of P. corethrurus increased the dissipation 4-fold. The endogeic earthworm P. corethrurus can thus be used to remediate hydrocarbon-contaminated soils in tropical regions.     

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.     

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.     

Accumulation of As,Pb, Zn,Cd and Cu and arbuscular mycorrhizal status in populations of Cynodon dactylon grown on metal-contaminated soils

Metal(loid) accumulation and arbuscular mycorrhizal (AM) status of the dominant plant species, Cynodon dactylon, growing at four multi-metal(loid)s-contaminated sites and an uncontaminated site of China were investigated. Up to 94.7 As mg kg^?1, 417 Pb mg kg^?1, 498 Zn mg kg^?1, 5.8 Cd mg kg^?1 and 27.7 Cu mg kg^?1 in shoots of C. dactylon were recorded. The plant was colonized consistently by AM fungi (33.0–65.5%) at both uncontaminated site and metal-contaminated sites. Based on morphological characteristics, fourteen species of AM fungi were identified in the rhizosphere of C. dactylon, with one belonging to the genus of Acaulospora and the other thirteen belonging to the genus of Glomus. Glomus etunicatum was the most common species associated with C. dactylon growing at metal-contaminated sites. Spore abundance in the rhizosphere of C. dactylon growing at the metal-contaminated soils (22–82 spores per 25 g soil) was significantly lower than that of the uncontaminated soils (371 spores per 25 g soil). However, AM fungal species diversity in the metal-contaminated soils was significantly higher than that in the uncontaminated soils. This is the first report of AM status in the rhizosphere of C. dactylon, the dominant plant survival in metal-contaminated soils. The investigation also suggests that phytorestoration of metal-contaminated sites might be facilitated using the appropriate plant with the aid of tolerant AM fungi.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

The combined effects of earthworms and arbuscular mycorrhizal fungi on microbial biomass and enzyme activities in a calcareous soil spiked with cadmium

Earthworms and arbuscular mycorrhizal fungi (AMF) are known to independently affect soil microbial and biochemical properties, in particular soil microbial biomass (SMB) and enzymes. However, less information is available about their interactive effects, particularly in soils contaminated with heavy metals such as cadmium (Cd). The amount of soil microbial biomass C (MBC), the rate of soil respiration (SRR) and the activities of urease and alkaline phosphatase (ALP) were measured in a calcareous soil artificially spiked with Cd (10 and 20 mg Cd kg⁻¹), inoculated with earthworm (Lumbricus rubellus L.), and AMF (Glomus intraradices and Glomus mosseae species) under maize (Zea mays L.) crop for 60 days. Results showed that the quantity of MBC, SRR and enzyme activities decreased with increasing Cd levels as a result of the elevated exchangeable Cd concentration. Earthworm addition increased soil exchangeable Cd levels, while AMF and their interaction with earthworms had no influence on this fraction of Cd. Earthworm activity resulted in no change in soil MBC, while inoculation with both AMF species significantly enhanced soil MBC contents. However, the presence of earthworms lowered soil MBC when inoculated with G. mosseae fungi, showing an interaction between the two organisms. Soil enzyme activities and SRR values tended to increase considerably with the inoculation of both earthworms and AMF. Nevertheless, earthworm activity did not affect ALP activity when inoculated with G. mosseae fungi, while the presence of earthworm enhanced urease activity only with G. intraradices species. The increases in enzyme activities and SRR were better ascribed to changes in soil organic carbon (OC), MBC and dissolved organic carbon (DOC) contents. In summary, results demonstrated that the influence of earthworms alone on Cd availability is more important than that of AMF in Cd-polluted soils; and that the interaction effects between these organisms on soil microorganism are much more important than on Cd availability. Thus, the presence of both earthworms and AMF could alleviate Cd effects on soil microbial life.     

Organic matter and polyacrylamide amendment of Norfolk loamy sand

Loamy sand soils of the southeastern USA Coastal Plains often have poor physical properties because they contain cemented subsurface hard layers that restrict root development and yield. Their physical properties can be improved by adding amendments. Polyacrylamide (PAM) amendments and/or organic matter (OM) in the form of ground wheat (Triticum aestivum L.) stubble or pecan (Carya illinoensis) branches were mixed into a blend of 90% E horizon and 10% Ap horizon (to assure microbial presence) obtained from a Norfolk soil (Acrisol or fine-loamy, siliceous, thermic Typic Kandiudult). We hypothesized that incorporation of these amendments would improve soil physical properties by reducing strength and improving aggregation. Amended treatments contained 450 g of soil, OM, and 30 or 120 mg kg⁻¹ of PAM (12 mg mol⁻¹, anionic, and 35% charge density); treatments were incubated for 96 days at 10% (w/w) water content. Twice during the incubation period, treatments were leached with 1.3 pore volumes of deionized water. After leaching and equilibrating to stable water contents, treatments were analyzed for bulk densities and probed with a 5-mm diameter flat-tipped bench-top penetrometer to measure penetration resistances. Though penetration resistances increased for the highest level of PAM amendment, they showed no significance when both PAM and OM were added to the soil. When compared to controls, treatments with PAM at 120 mg kg⁻¹ had decreased bulk densities. Treatments with both rates of PAM had decreased requirements for water needed to maintain treatments at 10% water contents. Aggregation increased with increasing amounts of PAM but showed no consistent trend when both PAM and OM were added to the soil. Because PAM increased aggregation and water holding capacities in these coastal soils, it could reduce the need for deep tillage. However, more work needs to be done to determine an effective mix of PAM and OM.     

Population size of indigenous Rhizobium leguminosarum biovar trifolii in long-term field experiments with sewage sludge cake,metal-amended liquid sludge or metal salts: Effects of zinc,copper and cadmium

There is conflicting evidence, and therefore continuing concern, as to whether metals in sewage sludge are deleterious to soil microbial processes and long-term agricultural productivity. Nine field experiments with sewage sludge cakes, three with metal-amended liquid sludges and three with inorganic metal salts were set up across Britain in 1994 to give individual metal dose–response treatments to try to answer this question. This study reports on the effects of Zn, Cu and Cd on the population size of Rhizobium leguminosarum biovar trifolii, a nitrogen fixing symbiont of white clover (Trifolium repens), in soils from these experiments over 11 years. Significant (P < 0.05) reductions in indigenous rhizobial numbers occurred on the Zn metal dose–response treatments at eight of the sludge cake sites in 2005, but few consistent effects were evident on the Cu or Cd metal dose–response treatments during the 11-year monitoring period. The soil total Zn concentrations where effects occurred were near to the UK statutory limit of 300 mg kg^?1 for soils receiving sewage sludge. No significant reductions occurred in any treatments on the metal-amended liquid sludge or inorganic metal salt experiments in which the metals would be expected to be in a more bioavailable form, even after 11 years. The effects in the sludge cake experiments were related consistently with soil total Zn, with no recovery to date. The reductions in clover rhizobial numbers in the sludge cake experiments were due to Zn effects on free-living rhizobia in the soil, with gradual die-off over a long time with increasing soil total Zn concentrations. Currently, no consistent adverse effects on rhizobia have been seen at the UK limits for Cu and Cd of 135 and 3 mg kg^?1, respectively.     

Influence of carbon amendments on soil denitrifier abundance in soil microcosms

It is known that carbon (C) amendments increase microbial activity in anoxic soil microcosm studies, however the effects on abundance of total and denitrifier bacterial communities is uncertain. Quantitative PCR was used to target the 16S rRNA gene for the total bacterial community, the nosZ functional gene to reflect a broad denitrifier community, and functional genes from narrow denitrifier communities represented by Pseudomonas mandelii and related species (cnorB_P) and Bosea/Bradyrhizobium/Ensifer spp. (cnorB_B). Repacked soil cores were amended with varying amounts of glucose and red clover plant tissue (0–1000 mg C kg^? 1 of soil) and incubated for 96 h. Carbon amendment significantly increased respiration as measured by cumulative CO₂ emissions. Inputs of red clover or glucose at 1000 mg C kg^? 1 of soil caused increased abundance in the total bacteria under the conditions used. There was about an approximate 2-fold increase in the abundance of bacteria bearing the nosZ gene, but only in treatments receiving 500 or 1000 mg C kg^? 1 of soil of glucose or red clover, respectively. Additions of ≥ 500 mg C kg^? 1 soil of red clover and ≥ 250 mg C kg^? 1 of glucose increased cnorB_P-gene bearing denitrifiers. Changes in abundance of the targeted communities were related to C availability in soil, as indicated by soil respiration, regardless of C source. Applications of C amendments at rates that would occur in agricultural soils not only increase microbial activity, but can also induce changes in abundance of total bacterial and denitrifier communities in studies of anoxic soil microcosms.     

The ecological dose value (ED50) for assessing Cd toxicity on ATP content and dehydrogenase and urease activities of soil

Two soils of contrasting texture, organic matter content and pH were treated with CdSO₄ solutions to give a Cd concentration range of 0–4000 mg kg⁻¹ soil. The content of ATP and dehydrogenase and urease activities of soils were assayed after 3 h, and 7 and 28 days of Cd contamination. The relative ED₅₀ values were calculated by two kinetic models (model 1 and model 2) used by Speir et al. (1995) and by the sigmoidal dose–response model (model 3) employed by Haanstra et al. (1985). Model 1 was the most successful in calculating the ED₅₀ values for the ATP content, urease and dehydrogenase activities when both soils were contaminated by Cd. Similar ED₅₀ values were predicted by model 1 (describing the full inhibition) and model 3 only when the correlation coefficients r² were higher than 0.9. The ED₅₀ values of ATP calculated by model 1 were markedly higher than those calculated by model 2 (describing partial inhibition) when both models gave correlation coefficients higher than 0.9. This behavior was due to the high asymptote values obtained using model 2. According to model 2, some of the enzyme activities responsible for the ATP synthesis were probably not inhibited at the highest Cd concentrations. The inhibitory effect of Cd on the ATP content and both enzymatic activities was lower in the Castelporziano soil, which had the highest total organic carbon content.     

Effects of nonylphenols on soil microbial activity and water retention

The main aim of this study is to analyze the influence of 4-nonylphenol (NP) on soil water retention and biological activity. Two doses of 4-nonylphenol (25 and 50 mg kg⁻¹) were tested in a loam soil with and without peat amendment. In general, one week after the start of the experiment, the soil water content retained at −0.75 MPa of soil suction was 18% higher in the soil amended and its basal respiration (BR) was 15% higher than soil without peat. In contrast, the microbial activity indices (CM: coefficient of mineralization or BR:total organic carbon (TOC) ratio; Cmic:Corg: microbial biomass carbon (MBC):TOC ratio; qCO₂: metabolic quotient or BR:MBC ratio) were higher in the soil without peat, compared to the soil amended with peat. On the other hand, the addition of NP to soil was able to modify soil biological but not physical (water retention, desorption) properties. When soil was amended with peat, MBC was reduced one week after applying NP. In contrast, no effects of NP on MBC were observed in the soil without peat. BR was reduced by 16% one week after applying 50 mg kg⁻¹ of NP to soil with peat, and was increased by 46% one week after applying 25 mg kg⁻¹ of NP to soil without peat. The effects of NP on MBC and BR could be associated more with the adsorption of NP by soil organic matter, while changes in CM or Cmic:Corg ratio were more closely related to changes in soil water retention. The potential toxic effects of NP (high qCO₂ values) were only observed in the absence of peat amendments. Peat addition reduced NP toxic effects on microorganisms.