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.     

Changes in enzyme activities and microbial biomass after “in situ” remediation of a heavy metal-contaminated soil

Microbial properties such as microbial biomass carbon (MBC), arylsulfatase, β-glucosidase and dehydrogenase activities, and microbial heterotrophic potential, together with several chemical properties such as pH, CaCl₂ soluble heavy metal concentrations, total organic carbon and hydrosoluble carbon were measured to evaluate changes in soil quality, after “in situ” remediation of a heavy metal-contaminated soil from the Aznalcóllar mine accident (Southern Spain, 1998). The experiment was carried out using containers, filled with soil from the affected area. Four organic amendments (a municipal waste compost, a biosolid compost, a leonardite and a litter) and an inorganic amendment (sugarbeet lime) were mixed with the top soil at the rate of 100 Mg ha⁻¹. Unamended soil was used as control. Agrostis stolonifera L. was sown in the containers. The soil was sampled twice: one month and six months after amendment application. In general, these amendments improved the soil chemical properties: soil pH, total organic carbon and hydrosoluble carbon increased in the amended soils, while soluble heavy metal concentrations diminished. At the same time, higher MBC, enzyme activities and maximum rate of glucose mineralization values were found in the organically amended soils. Plant cover was also important in restoring the soil chemical and microbial properties in all the soils, but mainly in those that were not amended organically. As a rule, remediation measures improved soil quality in the contaminated soils.     

长期施氮肥对黄棕壤微生物生物性状的影响及其调控因素

基于黄棕壤小麦-甘薯轮作的长期定位试验,探究不同施氮处理土壤微生物生物量碳(MBC)、氮(MBN)含量和酶活性的变化及其潜在调控因子,为科学施氮提高土壤质量和改善土壤生态功能提供依据。试验选取始于2011年4个施氮处理:不施肥(CK)、不施氮肥(PK)、施化学氮肥(NPK)和化学氮肥配施有机肥(NPKM),调查两季作物收获后土壤MBC和MBN含量、酶活性及微生物碳氮利用效率的变化,并通过冗余分析(RDA)和结构方程模型(SEM)明确调控弱酸性黄棕壤中MBC和MBN变化的潜在因素。小麦和甘薯两季的结果表明:施氮肥降低了土壤MBC、MBN含量和蔗糖酶(SSC)、脲酶(SUE)活性,与NPK处理相比, NPKM处理增加了MBC、MBN含量和SSC、SUE活性。长期施用氮肥提高了土壤有机碳(SOC)和土壤养分[全氮(TN)和矿质态氮(MN)]含量,但施氮肥显著降低土壤p H以及微生物的碳氮利用效率。与小麦季相比,甘薯季土壤SOC和MN含量有所下降,而MBN含量和SSC活性有所升高。RDA和SEM结果表明,氮肥的施用强化了MBC与MBN、SSC与MBC以及SUE与MBC之间的关联性;不同施氮处理下土壤p H、有机碳、氮含量以及微生物的碳氮利用效率的变化直接或间接地影响土壤MBC、MBN含量和SSC、SUE活性,其中p H是调控土壤MBC变化的直接因素,而土壤SSC和SUE活性与MBC、MBN含量相互影响。长期施用氮肥降低了黄棕壤MBC、MBN含量和酶活性,化学氮肥配施有机肥有利于缓解生物性状的下降,土壤p H是影响MBC变化的主要因素,小麦-甘薯轮作中土壤微生物强烈的碳代谢过程利于增加MBN。     

Soil microbial biomass and organic matter fractions during transition from conventional to organic farming systems

The aim of this study was to investigate the response of soil microbial biomass and organic matter fractions during the transition from conventional to organic farming in a tropical soil. Soil samples were collected from three different plots planted with Malpighia glaba: conventional plot with 10 years (CON); transitional plot with 2 years under organic farming system (TRA); organic plot with 5 years under organic farming system (ORG). A plot under native vegetation (NV) was used as a reference. Soil microbial biomass C (MBC) and N (MBN), soil organic carbon (SOC) and total N (TN), soil organic matter fractioning and microbial indices were evaluated in soil samples collected at 0–5, 5–10, 10–20 and 20–40 cm depth. SOC and fulvic acids fraction contents were higher in the ORG system at 0–5 cm and 5–10 cm depths. Soil MBC was highest in the ORG, in all depths, than in others plots. Soil MBN was similar between ORG, TRA and NV in the surface layer. The lowest values for soil MBC and MBN were observed in CON plot. Soil microbial biomass increased gradually from conventional to organic farming, leading to consistent and distinct differences from the conventional control by the end of the second year.     

Effects of vegetation on soil microbial C,N, and P dynamics in a tropical forest and savanna of Central Africa

The forest–savanna transition zone is widely distributed on nutrient-poor oxisols in Central Africa. To reveal and compare the nutrient cycle in relation to soil microbes for forest and savanna vegetation in this area, we evaluated seasonal fluctuations in microbial biomass carbon (MBC), nitrogen (MBN), and phosphorus (MBP) for 13 months as well as soil moisture, temperature, soil pH levels, and nutrients for both vegetation types in eastern Cameroon. Soil pH was significantly lower in forest (4.3) than in savanna (5.6), and soil N availability was greater in forest (87.1 mg N kg⁻¹ soil) than in savanna (32.9 mg N kg⁻¹ soil). We found a significant positive correlation between soil moisture and MBP in forest, indicating the importance of organic P mineralization for MBP, whereas in savanna, we found a significant positive correlation between soil N availability and MBP, indicating N limitation for MBP. These results suggest that for soil microbes, forest is an N-saturated and P-limited ecosystem, whereas savanna is an N-limited ecosystem. Additionally, we observed a significantly lower MBN and larger MB C:N ratio in forest (50.7 mg N kg⁻¹ soil and 8.6, respectively) than in savanna (60.0 mg N kg⁻¹ soil and 6.5, respectively) during the experimental period, despite the rich soil N condition in forest. This may be due to the significantly lower soil pH in forest, which influences the different soil microbial communities (fungi-to-bacteria ratio) in forest versus savanna, and therefore, our results indicate that, in terms of microbial N dynamics, soil pH rather than soil substrate conditions controls the soil microbial communities in this area. Further studies should be focused on soil microbial community, such as PLFA, which was not evaluated in the present study.     

The relationships among microbial parameters and the rate of organic matter mineralization in forest soils,as influenced by forest type

Vegetation type influences the rate of accumulation and mineralization of organic matter in forest soil, mainly through its effect on soil microorganisms. We investigated the relationships among forest types and microbial biomass C (MBC), basal respiration (R_B), substrate-induced respiration (R_S), N mineralization (N_min), specific growth rate μ, microbial eco-physiology and activities of seven hydrolytic enzymes, in samples taken from 25 stands on acidic soils and one stand on limestone, covering typical types of coniferous and deciduous forests in Central Europe. Soils under deciduous trees were less acidic than soils of coniferous forests, which led to increased mineralizing activities R_B and N_min, and a higher proportion of active microbial biomass (R_S/MBC) in the Of horizon. This resulted in more extractable organic C (0.5 M K₂SO₄) in soils of deciduous forests and a higher accumulation of soil organic matter (SOM) in coniferous forest soil. No effect of forest type on the microbial properties was detected in the Oh horizon and in the 0–10 cm layer. The microbial quotient (MBC/C_org), reflecting the quality of organic matter used for microbial growth, was higher in deciduous forests in all three layers. The metabolic quotient qCO₂ (R_B/MBC) and the specific growth rate μ, estimated using respiration growth curves, did not differ in soils of both forest types. Our results showed that the quality of SOM in coniferous forests supported microorganisms with higher activities of β-glucosidase, cellobiosidase and β-xylosidase, which suggested the key importance of fungi in these soils. Processes mediated by bacteria were probably more important in deciduous forest soils with higher activities of arylsulphatase and urease. The results from the stand on limestone showed that pH had a positive effect on microbial biomass and SOM mineralization.     

Biochar soil amendment increased bacterial but decreased fungal gene abundance with shifts in community structure in a slightly acid rice paddy from Southwest China

Biochar’s role on greenhouse gas emission and plant growth has been well addressed. However, there have been few studies on changes in soil microbial community and activities with biochar soil amendment (BSA) in croplands. In a field experiment, biochar was amended at rates of 0, 20 and 40 t ha⁻¹ (C0, C1 and C2, respectively) in May 2010 before rice transplantation in a rice paddy from Sichuan, China. Topsoil (0–15 cm) was collected from the rice paddy while rice harvest in late October 2011. Soil physico-chemical properties and microbial biomass carbon (MBC) and nitrogen (MBN) as well as selected soil enzyme activities were determined. Based on 16S rRNA and 18S rRNA gene, bacterial and fungal community structure and abundance were characterized using terminal-restriction fragment length polymorphism (T-RFLP) combined with clone library analysis, denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR assay (qPCR). Contents of SOC and total N and soil pH were increased but bulk density decreased significantly. While no changes in MBC and MBN, gene copy numbers of bacterial 16S rRNA was shown significantly increased by 28% and 64% and that of fungal 18S rRNA significantly decreased by 35% and 46% under BSA at 20 and 40 t ha⁻¹ respectively over control. Moreover, there was a significant decrease by 70% in abundance of Methylophilaceae and of Hydrogenophilaceae with an increase by 45% in Anaerolineae abundance under BSA at 40 t ha⁻¹ over control. Whereas, using sequencing DGGE bands of fungal 18S rRNA gene, some bands affiliated with Ascomycota and Glomeromycota were shown inhibited by BSA at rate of 40 t ha⁻¹. Significant increases in activities of dehydrogenase, alkaline phosphatases while decreased β-glucosidase were also observed under BSA. The results here indicated a shift toward a bacterial dominated microbial community in the rice paddy with BSA.     

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.     

Species richness and phylogenetic diversity comparisons of soil microbial communities affected by nickel-mining and revegetation efforts in New Caledonia

In order to understand the impact of human activities on soil microbial diversity, we investigated bacterial communities in samples recovered from four New Caledonia environments that have been disturbed by varying degrees of nickel mining associated activities: an undisturbed area with natural soil (characterized by pristine vegetation), a mine spoil (devoid of vegetation), two revegetated mine spoils by endemic plants. For each sample, total DNA was extracted and 16S rDNA clone library were constructed. 442 clones were sequenced and analyzed. Using these clones, diversity was estimated not only in terms of species richness (non-parametric estimators) and evenness (Reciprocal of Simpson's index), but also in terms of phylogenetic diversity (LIBSHUFF program). Statistically significant differences were detected in phylogenetic composition between mine spoils and natural soil (p = 0.001), between revegetated soils and natural soil (p = 0.001), and between revegetated soils and mine spoils (p = 0.001). On the other hand, no significant differences in species richness were observed between the different environmental samples.These findings provide insights into the response of bacterial community following environmental perturbations caused by nickel-mining activities and revegetation efforts.     

Effects of long-term sewage irrigation on agricultural soil microbial structural and functional characterizations in Shandong,China

Soil samples taken from a sewage irrigation area, a partial sewage irrigation area and a ground water irrigation area (control area) were studied with the methods of Biolog and FAME. It was found that the microbial utilization of carbon sources in sewage irrigation areas was much higher than that of control area (P < 0.05). With the increasing of the amount of sewage irrigation, microbial functional diversity slightly increased by the Biolog analysis; however, the amount of epiphyte decreased by the FAME analysis. The results also showed that the Cr, Zn contents were positively correlated with the values of AWCD and the microbial diversity, while Hg content showed negative correlation with the microbial parameters (AWCD of 72 h and Shannon index). Our studies suggested that sewage irrigation resulted in an obvious increase of heavy metals content in soil (P < 0.05), although the maximum heavy metals concentrations were much lower than the current standard of China. Other soil basic characteristics such as cation exchange capacity (CEC), total nitrogen (N_t) and organic matter in sewage irrigation areas obviously increased (P < 0.05). Therefore, it is demonstrated that long-term sewage irrigation had influenced soil microorganisms and soil quality in the studied soils. As a result, it is important to monitor the changes in agricultural soils. Furthermore, our results also confirmed that the methods of Biolog and FAME are effective tools for the assessment of soil microbial structure/function and soil health.     

Animal manure and anhydrous ammonia amendment alter microbial carbon use efficiency,microbial biomass,and activities of dehydrogenase and amidohydrolases in semiarid agroecosystems

The potential negative impact of agricultural practices on soil and water quality is of environmental concern. The associated nutrient transformations and movements that lead to environmental concerns are inseparable from microbial and biochemical activities. Therefore, biochemical and microbiological parameters directing nitrogen (N) transformations in soils amended with different animal manures or inorganic N fertilizers were investigated. Soils under continuous corn cultivation were treated with N annually for 5 years at 56, 168, and 504 kg N ha⁻¹ in the form of swine effluent, beef manure, or anhydrous ammonia. Animal manure treatments increased dehydrogenase activity, microbial biomass carbon (C_mic) and N (N_mic) contents, and activities of amidohydrolases, including l-asparaginase, urease, l-glutaminase, amidase, and β-glucosaminidase. Soils receiving anhydrous ammonia demonstrated increased nitrate contents, but reduced microbiological and biochemical activities. All treatments decreased C_mic:organic C (C_org) ratios compared with the control, indicating reduced microbial C use efficiency and disturbance of C equilibrium in these soil environments. Activities of all enzymes tested were significantly correlated with soil C_org contents (P < 0.001, n = 108), but little correlation (r = 0.03, n = 36) was detected between C_mic and C_org. Activities of amidase and β-glucosaminidase were dominated by accumulated enzymes that were free of microbial cells, while activities of asparaginase and glutaminase were originated predominately from intracellular enzymes. Results indicated that soil microbial and biochemical activities are sensitive indicators of processes involved in N flow and C use efficiency in semiarid agroecosystems.     

不同耕作方式对土壤有机碳、微生物量及酶活性的影响

【目的】依托8年长期(2005~2012)固定道定位试验,研究不同耕作方式对土壤有机碳、土壤微生物量、土壤酶活性在0—90 cm土层的分布特征,为优化中国西北干旱区的耕作方式提供理论依据。【方法】试验包括固定道垄作(PRB)、固定道平作(PFT)与传统耕作(CT)三种耕作模式下的土壤有机碳土壤总有机碳(TOC)、颗粒有机碳(POC)、土壤微生物量碳(MBC)、土壤微生物量氮(MBN)、土壤微生物量磷(MBP)、蔗糖酶、过氧化氢酶、脲酶及小麦产量进行了测定和分析。【结果】在0—90 cm土层,不同耕作方式下的TOC、POC、MBC、MBN、MBP、蔗糖酶活性、脲酶活性均随着土层的增加呈下降趋势,过氧化氢酶活性呈先下降后增大的分布特征;在0—60 cm,固定道保护性耕作能够显著增加心土层作物生长带土壤有机碳储量,有机碳储量大小为PRBPFTCT;PRB、PFT较CT可以显著增加0—10 cm作物生长带TOC、POC、MBC、MBN、MBP含量、蔗糖酶、脲酶活性,其大小为PRBPFTCT;耕作方式对过氧化氢酶活性影响不显著;TOC、POC、MBC、MBN、MBP、蔗糖酶活性、脲酶活性、过氧化氢酶活性之间均达到了显著或极显著相关。【结论】PRB较PFT、CT能够提高耕作层(0—10 cm)土壤有机碳含量、土壤微生物量、土壤酶活性, 增加作物产量, 增大0—60 cm土层有机碳储量,耕作方式(PRB、PFT及CT)对10 cm以下土层土壤环境改善作用不明显。     

Changes in soil microbial biomass and functional diversity with a nitrogen gradient in soil columns

Fertilization generates nutrient patches that may impact soil microbial activity. In this study, nitrogen patches were generated by adding ammonium sulfate or urea to soil columns (length 25 cm; internal diameter 7.2 cm). Changes in nitrogen transformation, soil microbial biomass, and microbial functional diversity with the nitrogen gradients were investigated to evaluate the response of microbial activity to chemical fertilizer nutrient patches. After applying of ammonium sulfate or urea, the added nitrogen migrated about 7 cm. Microbial biomass carbon (MBC) was lower in fertilized soil than in the control (CK) treatment at the same soil layers. MBC increased with soil depth while microbial biomass nitrogen (MBN) decreased. BIOLOG analysis indicated that the average well color development (AWCD) and functional diversity indices of the microbial communities were lower in the 1 cm and 2 cm soil layers after application of ammonium sulfate; the highest values were in the 3 cm soil layer. AWCD and Shannon indices from the 1 to 5 cm soil layers were higher than those from other soil layers under urea application. Both principal component analysis and carbon substrate utilization analysis showed significant separation of soil microbial communities among different soil layers under application of ammonium sulfate or urea. Microbial activity was substantially decreased when NH₄⁺-N concentration was higher than 528.5 mg kg⁻¹ (1–3 cm soil layer under ammonium sulfate application) or 536.8 mg kg⁻¹ (1 cm soil layer under urea application). These findings indicated that changes in soil microbial biomass and microbial functional diversity can occur with a nitrogen gradient. The extent of changes depends on the nitrogen concentration and the form of inorganic fertilizer.     

Complementarity of bioassays and microbial activity measurements for the evaluation of hydrocarbon-contaminated soils quality

In order to describe a soil polluted with hydrocarbons, the complementarity of bioassays and microbial activities measurements was studied. The samples of soil were taken from a site which had received oil tank residues over 50 years. Five zones were sampled. Each sample was characterized by chemical analyses, the measurement of dehydrogenase, phosphatase, hydrolysis of FDA and urease activities, soil respiration, and Microtox and Metplate bioassays. The chemical analyses revealed different levels of total hydrocarbon concentrations (from 1.5 to 78.8 mg/kg of dry soil) but also relatively high quantities of nickel (from 14.5 to 841.6 mg/kg of dry soil) and lead (30.9–355.4 mg/kg of dry soil) or cadmium (0–1.2 mg/kg of dry soil) in the different zones. Urease and dehydrogenase were sensitive to the presence of metals (31% inhibition of urease and 50% inhibition of dehydrogenase in the most contaminated soil). Measurements of Substrate Induced Respiration showed that the soil microflora were stressed in the presence of the pollutants. In the zone containing the highest concentration of metals, the microbial activities were low and the bioassays revealed a high potential toxicity (e.g. IC50 for Microtox obtained with a 15% dilution of soil, 90% inhibition of β-galactosidase activity). In the other zones, the soil microbial activities were not depressed in comparison to the reference zone whereas the bioassays revealed the presence of toxic compounds extracted with the solvent used.     

Rhizosphere microbial community and Zn uptake by willow (Salix purpurea L.) depend on soil sulfur concentrations in metalliferous peat soils

On numerous occasions, rhizosphere microbial activities have been identified as a key factor in metal phytoavailability to various plant species and in phytoremediation of metal-contaminated sites. For soil bioremediation efforts in heavy metal contaminated areas, microbes adapted to higher concentrations of heavy metals are required. This study was a field survey undertaken to examine rhizosphere microbial communities and biogeochemistry of soils associated with Zn accumulation by indigenous willows (Salix purpurea L.) in the naturally metalliferous peat soils located near Elba, NY. Soil and willow leaf samples were collected from seven points, at intervals 18 m apart along a willow hedgerow, on four different dates during the growing season. Soil bacterial community composition was characterized by terminal restriction fragment length polymorphism (T-RFLP) analysis and a 16S clone library was created from the rhizosphere of willows and soils containing the highest concentrations of Zn. Bacterial community composition was correlated with soil sulfate, but not with soil pH. The clone library revealed comparable phylogenetic associations to those found in other heavy metal-contaminated soils, and was dominated by affiliations within the phyla Acidobacteria (32%), and Proteobacteria (37%), and the remaining clones were associated with a wide array of phyla including Actinobacteria, Gemmatimonadetes, Planctomycetes, Verrucomicrobia, Bacteriodetes, and Cyanobacteria. Diverse microbial populations were present in both rhizosphere and bulk soils of these naturally metalliferous peat soils with community composition highly correlated to the soil sulfate cycle throughout the growing season indicative of a sulfur-oxidizing rhizosphere microbial community. Results confirm the importance of soil characterization for informing bioremediation efforts in heavy metal contaminated areas and the reciprocity that microbial communities uniquely adapted to specific conditions and heavy metals may have on an ecosystem.     

Effects of arbuscular mycorrhizal inoculation on the growth of Elsholtzia splendens and Zea mays and the activities of phosphatase and urease in a multi-metal-contaminated soil under unsterilized conditions

A pot culture experiment was carried out to study the effects of arbuscular mycorrhizal (AM) inoculation on the growth of Elsholtzia splendens and Zea mays and the activities of phosphatase and urease in a soil contaminated with Cu, Zn, Pb and Cd. Two AM fungal inocula, MI containing Glomus caledonium and MII containing Gigaspora margarita,Gigaspora decipens, Scutellospora gilmori, Acaulospora spp. and Glomus spp., were applied to the soil. The plants of E. splendens and Z. mays were harvested after 24 and 10 weeks of growth, respectively. Both plant species had a similar trend in mycorrhizal colonization rates, MI > MII > control. Shoot and root biomass of Z. mays was increased by MI, while not affected significantly by MII. Although both MI and MII increased plant dry weight of E. splendens, MII was more effective. Mycorrhizal dependency (MD) with MI and MII was 14.8 and 33.5, respectively for E. splendens, and 11.0 and 0.9, respectively for Z. mays. Both inocula increased the activities of phosphatase and urease in the soils of E. splendens and Z. mays, but MI was more effective than MII for urease, while MII more effective than MI for phosphatase. Although the mechanisms involved in these responses are not clear, AM fungal inoculum may be important and used for the phytoremediation of heavy metal contaminated soils, but both inoculum type and host species must be considered.     

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.     

C and N mineralization and microbial biomass in heavy-metal contaminated soil

Heavy metals such as arsenic (As), lead (Pb), copper (Cu) and zinc (Zn) can be found in large concentrations in mine spills in Mexico. Interest in contamination by these heavy metals has increased recently as they can change the functioning of soil ecosystems qualitatively and quantitatively. They disturb the activities of soil fauna and contaminate drinking water in large parts of the world, which severely affects human health. Little, however, is known how heavy metals might affect the biological functioning of a soil. Soil was sampled from eight locations along a gradient of heavy-metal contamination with distance from a mine in San Luis Potosí (Mexico) active since about 1800 AD. Microbial biomass was determined with the original chloroform fumigation incubation (CFI) as well as extraction (CFE) techniques and the substrate induced respiration (SIR) technique while C and N mineralization were measured. Total concentrations of As in the top 0–10 cm soil layer ranged from 8 to 22992 mg kg^–1, from 31 to 1845 mg kg^–1 for Pb, from 27 to 1620 mg kg^–1 for Cu and from 81 to 4218 mg kg^–1 for Zn. There was a significant negative correlation (P < 0.0001) between microbial biomass, soil organic carbon, total N and C mineralization and the heavy metal content of the soil. The microbial biomass C to organic C ratio, which varied from 0.4 to 1.9%, specific respiratory activity (qCO₂), and oxidation of NO₂^– were not affected by heavy metals. It was found that long-term contamination of soil with heavy metals had an adverse effect on the amount of soil microorganisms as evidenced by a marked decrease in microbial biomass C, but not some of their characteristics. According to principal components analysis (PCA), the correlation matrix showed three distinct factors explaining 71% of the variance. A first factor including heavy metals (As, Pb, Cu and Zn) with a negative loading and total N, organic C, soil microbial biomass with a positive loading characterized the soil organic matter and contamination status. Loam and sand combined for the second factor characterizing the textural classification while the third factor was loaded by CEC and clay content.     

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.     

Relative strengths of relationships between plant,microbial, and environmental parameters in heavy-metal contaminated floodplain soil

We used a combination of sampling and statistical approaches to investigate the relative influence of metals, soil acidity, and organic matter on a suite of analogous plant and microbial community parameters in floodplain soils contaminated by mine wastes in the early twentieth century. We compared the sensitivity of plant and microbial communities to environmental variables and to one another using constrained ordination analyses. Environmental factors accounted for a larger percentage of the total variance in microbial communities (56.2%) than plant communities (22.0%). We also investigated biological and geochemical changes that occurred along a short transect (64 cm) that spanned a transition from productive grassland to an area of barren wasteland representing a total functional collapse of the grassland/soil ecosystem. Along this small-scale transect we quantified geochemical parameters and biological parameters in two soil layers, an upper layer (0–10 cm) and a lower layer (10–20 cm). Results from the short transect indicated that soil respiration was not a strong indicator of underlying metal concentrations, but soil acidity was correlated in the upper and lower layers. PLFA profiles changed with distance along the gradient in the upper, but not the lower layer. Implications for remediation of contaminated floodplain soils are discussed.