Sewage sludge processing determines its impact on soil microbial community structure and function

Composting and thermal drying are amongst the most commonly used post-digestion processes for allowing sanitation and biological stabilization of sewage sludge from municipal treatment plants, and making it suitable as soil conditioner for use in agriculture. To assess the impact of sludge-derived materials on soil microbial properties, fresh (LAF), composted (LAC) and thermally dried (LAT) sludge fractions, each resulting from a different post-treatment process of a same aerobically digested sewage sludge, were added at 1% (w/w) application rate on two contrasting (a loam and a loamy sand) soils and incubated under laboratory conditions for 28 days. Soil respiration, microbial ATP content, hydrolytic activities and arginine ammonification rate were monitored throughout the incubation period. Results showed that soil biochemical variables, including the metabolic quotient (qCO₂), were markedly stimulated after sludge application, and the magnitude of this stimulatory effect was dependent on sludge type (precisely LAT > LAF > LAC), but not on soil type. This effect was related to the content of stable organic matter, which was lower in LAT. Genetic fingerprinting by PCR–DGGE revealed that compositional shifts of soil bacterial and, at greater extent, actinobacterial communities were responsive to the amendment with a differing sludge fraction. The observed time-dependent changes in the DGGE profiles of amended soils reflected the microbial turnover dependent on the sludge nutrient input, whereas no indications of adverse effects of sludge-borne contaminants were noted. Our findings indicate that composting rather thermal drying can represent a more appropriate post-digestion process to make sewage sludge suitable for use as soil conditioner in agriculture.     

Influences of non-herbaceous biochar on arbuscular mycorrhizal fungal abundances in roots and soils: Results from growth-chamber and field experiments

Biochar holds promise as an amendment for soil quality improvement and sequestration of atmospheric carbon dioxide. However, knowledge of how biochar influences soil properties, especially soil microorganisms, is limited. Three separate studies were conducted, with two studies using Plantago lanceolata as the AMF hosting plant, and a third being conducted in the field. Each of the three studies employed a different soil type. Furthermore, a total of five different biochars, and ten different biochar application rates, were used across the three experiments. All experiments had the goal to examine biochar influences on arbuscular mycorrhizal fungal (AMF) abundance in roots and AMF abundance (hyphal lengths) in soils. AMF abundance was either decreased or remained unchanged across all biochar treatments. When AMF abundances decreased, significant changes in soil properties, primarily in soil P availability, were observed. Application of large quantities (2.0% and 4.0%, w/w) of a lodgepole pine biochar, led to significant declines in AMF abundance in roots of 58% and 73% respectively, but not in soils. These declines in AMF abundance were accompanied by significant declines (28% and 34%) in soil P availability. After addition of a peanut shell biochar produced at 360 °C, P increased by 101% while AMF root colonization and extraradical hyphal lengths deceased by 74% and 95% respectively. Field application of mango wood biochar at rates of 23.2 and 116.1 t C ha⁻¹ increased P availabilities by 163% and 208% respectively and decreased AMF abundances in soils by 43% and 77%. These findings may have implications for soil management where the goal is to increase the services provided by AMF.     

Microbial mineralization of biochar and wheat straw mixture in soil: A short-term study

A short-term incubation study was carried out to investigate the effect of biochar addition to soil on CO₂ emissions, microbial biomass, soil soluble carbon (C) nitrogen (N) and nitrate–nitrogen (NO₃–N). Four soil treatments were investigated: soil only (control); soil + 5% biochar; soil + 0.5% wheat straw; soil + 5% biochar + 0.5% wheat straw. The biochar used was obtained from hardwood by pyrolysis at 500 °C. Periodic measurements of soil respiration, microbial biomass, soluble organic C, N and NO₃–N were performed throughout the experiment (84 days). Only 2.8% of the added biochar C was respired, whereas 56% of the added wheat straw C was decomposed. Total net CO₂ emitted by soil respiration suggested that wheat straw had no priming effect on biochar C decomposition. Moreover, wheat straw significantly increased microbial C and N and at the same time decreased soluble organic N. On the other hand, biochar did not influence microbial biomass nor soluble organic N. Thus it is possible to conclude that biochar was a very stable C source and could be an efficient, long-term strategy to sequester C in soils. Moreover, the addition of crop residues together with biochar could actively reduce the soil N leaching potential by means of N immobilization.     

Bacterial community structure and microbial activity in different soils amended with biogas residues and cattle slurry

Anaerobic digestion of organic materials generates residues of differing chemical composition compared to undigested animal manures, which may affect the soil microbial ecosystem differently when used as fertilizers. This study investigated the effects of two biogas residues (BR-A and BR-B) and cattle slurry (CS) applied at rates corresponding to 70 kg NH₄⁺-N ha⁻¹ on bacterial community structure and microbial activity in three soils of different texture (a sandy, a clay and an organic clay soil). 16S rRNA genes were targeted in PCR reactions and bacterial community profiles visualized using terminal restriction fragment length polymorphism. General microbial activity was measured as basal respiration (B-resp), substrate-induced respiration (SIR), specific growth rate (μ_SIR), metabolic quotient (qCO₂) and nitrogen mineralization capacity (NMC). Non-metric multidimensional scaling analysis visualized shifts in bacterial community structure related to microbial functions. There were significant differences in bacterial community structure after 120 days of incubation (+20 °C at 70% of WHC) between non-amended (control) and amended soils, especially in the sandy soil, where CS caused a more pronounced shift than biogas residues. Terminal-restriction fragment (TRF) 307, the predominant peak in CS-amended sandy soil, was identified as possibly Bacillus or Streptococcus. TRF 226, the dominant peak in organic soil amended with BR-B, was classified as Rhodopseudomonas. B-resp significantly increased and SIR decreased in all amendments to organic soil compared with the control, potentially indicating decreased efficiency of heterotrophic microorganisms to convert organic carbon into microbial biomass. This was also reflected in an elevated qCO₂ in the organic soil. The μ_SIR level was higher in the sandy soil amended with BR-A than with BR-B or CS, indicating a shift toward species capable of rapidly utilizing glucose. NMC was significantly elevated in the clay and organic soils amended with BR-A and BR-B and in the sandy soil amended with BR-B and CS. Thus, biogas residues and cattle slurry had different effects on the bacterial community structure and microbial activity in the three soils. However, the effects of biogas residues on microbial activities were comparable in magnitude to those of cattle slurry and the bacterial community structure was less affected. Therefore, we do not see any reason not to recommend using biogas residues as fertilizers based on the results presented.     

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.     

Evaluation of pinewood biochar as a carrier of bacterial strain Enterobacter cloacae UW5 for soil inoculation

The large-scale production of biochar for carbon sequestration provides an opportunity for using these materials as inoculum carriers to deliver plant growth-promoting rhizobacteria (PGPR) into agricultural soils. Here, we evaluated the suitability of a biochar produced from pinewood pyrolyzed at 300 °C as a carrier for a well-studied PGPR strain, Enterobacter cloacae UW5. This strain was genetically modified to produce a green fluorescent protein marker that enabled tracking of the inoculum. Results from selective plate count assays and quantitative PCR (qPCR) confirmed that cell survival was slightly improved by addition of bacteria to soil using biochar as a carrier for the inoculant, as compared to soil directly inoculated. Total 16S rRNA genes were quantified using qPCR and DNA templates from the same soil treatments to distinguish the impact of biochar on total bacterial abundance from its influence on inoculum survival. Here total bacterial abundance was not influenced by biochar. All treatments resulted in bacterial colonization of roots at population densities of approximately 10⁵ CFU g⁻¹ root mass. Cucumber plants grown in the biochar amended soils had significantly greater biomass and root development than those planted in un-amended soil, regardless of the presence of inoculum. The ability of bacteria to colonize the plant roots and produce a plant growth hormone was not affected by biochar. However, UW5 inoculum did not promote root development in cucumber in any of the soils tested here. Overall, these experiments suggest that the 300 °C pine biochar is effective for evenly distributing inoculum into soil and promotes cucumber development in sandy loams.     

Role of amendments on N cycling in Mediterranean abandoned semiarid soils

Soils found in semiarid areas of the Mediterranean Basin are particularly prone to degradation due to adverse climatic conditions with annual rainfall <300 mm and high temperatures being responsible for the scant vegetal growth and the consequent lack of organic matter. A three-year field experiment was conducted to test the potential of two organic amendments (sludge and compost) to improve soil quality and plant growth in a semiarid degraded Mediterranean ecosystem. Since little is known about N dynamics in such assisted ecosystems, we investigated the effects of this practice on key processes of the global N cycle. Besides soil chemical and biological parameters and vegetation cover, we measured absolute and specific potential nitrification and denitrification rates and quantified the size of the ammonia oxidising and denitrifying bacterial populations via quantitative PCR (amoA and nirS genes). At the end of the experiment soil fertility, microbial activity and plant growth had improved in treated plots. Amendments increased the amount of ammonia oxidisers and denitrifiers in soil, but the relative proportion of these groups varied in relation to the total microbial community, being higher in the case of ammonia oxidisers but not in the case of denitrifiers. As a consequence, significantly higher potential nitrification and denitrification rates were measured on a global basis in amended soils. Yet specific activities (potential rate/gene copy numbers) were lower for ammonia oxidisers in amended soils and for denitrifiers in sludge treated soils than those observed in control plots. Organic amendments influenced resource availability, the size and the activity patterns of microbial populations involved in long-term N dynamics. Therefore N cycling processes may play a key role to assist sustainable restoration practices in semiarid degraded areas.     

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.     

Soil microbial biomass,crop yields,and bacterial community structure as affected by long-term fertilizer treatments under wheat-rice cropping

Soil microbial biomass carbon (SMBC) and nitrogen (SMBN), soil microbial community structure, and crop yields were studied in a long-term (1982–2004) fertilization experiment carried out in Suining, Sichuan province of PR China. Eight treatments included three chemical fertilizer (CF) treatments (N, NP, NPK), three CF + farmyard manure (M) treatments (NM, NPM, NPKM), M alone and no fertilizer (CK) as control. The results showed that the soil microbial biomass was higher in soil treated with CFM than in soil treated with CF alone, and that NPKM gave the highest rice and wheat yields. The SMBC and SMBN were higher after rice than those after wheat cropping. SMBC correlated closely with soil organic matter. Average yields of wheat and rice for 22 years were higher and more stable in the fertilized plots than in control plots. Bacterial community structure was analyzed by PCR-DGGE targeting eubacterial 16S rRNA genes. A higher diversity of the soil bacterial community was found in soil amended with CFM than in other fertilizer treatments. Some specific band emerged in the soil amended with M. The highest diversity of bacterial communities was found in the NPKM treated soil. The bacterial community structures differed in rice and wheat plots. Sequencing of PCR products separated in DGGE showed that some of the common and dominant bands were closely related to Aquicella lusitana and to Acidobacteria. This study demonstrated that mixed application of N, P, and K with additional M amendment increased soil microbial biomass, diversified the bacterial communities and maintained the crop production in the Calcareous Purplish Paddy soil.     

Short-term mesofauna responses to soil additions of corn stover biochar and the role of microbial biomass

Biochar additions have been suggested to influence soil microbial communities that, through a cascade effect, may also impact soil fauna. In turn, any direct biochar effects on fauna can influence microbial communities through grazing, physical fragmentation of organic debris (and biochar) and modifying soil structure. If biochar creates a favorable environment for soil microorganisms, it is also plausible for fauna to be attracted to such microbially enriched habitats. However, how soil fauna respond to biochar addition to soil and what are the main factors that drive their behavior has rarely been experimentally addressed. Therefore, the behavior of two mesofauna species was assessed as a result of corn stover biochar (slow pyrolysis at 600 °C) additions to a loamy temperate soil, after preincubation for 2, 17, 31 and 61 d, and related to variations in microbial biomass and activity. Microbial biomass increased by 5–56% and activity by 6–156% with increasing biochar rates for the different preincubation times. Over the incubation time, microbial biomass did not change or increased at most 15% with the different biochar rates, while in turn microbial activity decreased steadily (around 70–80% at day 61). Enchytraeids generally did not show avoidance or preference to biochar when provided with an alternative unamended soil, while collembolans often showed avoidance responses. However, collembolan avoidance to biochar decreased or disappeared in biochar mixtures with higher microbial biomass and water extractable NH₄-N content, agreeing with the plausible role of microorganisms to potentially attract soil fauna after biochar applications. Avoidance response was mainly explained by environmental preferences of the test species and not by any toxic effect of the biochar in this study. However, avoidance after the application of biochar may still need to be considered due to the potential negative impacts of individuals’ migration on soil ecosystem functioning.     

Improving soil structure by promoting fungal abundance with organic soil amendments

Building soil structure in agroecosystems is important because it governs soil functions such as air and water movement, soil C stabilization, nutrient availability, and root system development. This study examined, under laboratory conditions, effects of organic amendments comprised of differing proportions of labile and semi-labile C on microbial community structure and macroaggregate formation in three variously textured soils where native structure was destroyed. Three amendment treatments were imposed (in order of increasing C lability): vegetable compost, dairy manure, hairy vetch (Vicia villosa Roth). Formation of water stable macroaggregates and changes in microbial community structure were evaluated over 82 days. Regardless of soil type, formation of large macroaggregates (LMA, >2000 μm diameter) was highest in soils amended with vetch, followed by manure, non-amended control, and compost. Vetch and manure had greater microbially available C and caused an increase in fungal biomarkers in all soils. Regression analysis indicated that LMA formation was most strongly related to the relative abundance of the fungal fatty acid methyl ester (FAME) 18:2ω6c (r = 0.55, p < 0.001), fungal ergosterol (r = 0.58, p < 0.001), and microbial biomass (r = 0.57, p < 0.001). Non-metric multidimensional scaling (NMS) ordination of FAME profiles revealed that vetch and manure drove shifts toward fungal-dominated soil microbial communities and greater LMA formation in these soils. This study demonstrated that, due to their greater amounts of microbially available C, vetch or manure inputs can be used to promote fungal proliferation in order to maintain or improve soil structure.     

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.     

Identifying indicators of C and N cycling in a clayey Ultisol under different tillage and uses in winter

Although tropical and subtropical environments permit two cropping cycles per year, maintaining adequate mulching on the soil surface remains a challenge. In some cases, leaving soils fallow during the winter as an agricultural practice to control pathogens contributes to reduce soil mulching. The aim of this study was to assess attributes associated with C and N cycling in a soil under conventional and no-tillage management, with contrasting uses in winter: black oats (Avena strigosa Schreb) as cover crop or fallow. No-tillage increased total C and N, irrespective the winter crop. Cropping black oats under no-tillage resulted in more microbial biomass C and N, and glutaminase activity (15.2%, 65.2%, and 24%, respectively) than no-tillage under fallow. Under conventional tillage, winter cropping did not affect the attributes under study. Available P was higher in the no-tillage system (9.2–12.3 mg kg⁻¹), especially when cropped with black oats, than in the conventional tillage system (4.8–6.6 mg kg⁻¹). A multivariate analysis showed strong relationships between soil microbiological and chemical attributes in the no-tillage system, especially when cropped with black oats. Soil pH, dehydrogenase and acid phosphatase activities were the most effective at separating the soil use in winter. Microbial N, total N, microbial to total N ratio, available P, metabolic quotient (qCO₂), and glutaminase activity were more effective at separating soil management regimes. The no-tillage system in association with winter oat cropping stimulated the soil microbial community, carbon and nutrient cycling, thereby helping to improve the sustainability of the cropping system.     

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.     

Lumbricus terrestris L. does not impact on the remediation efficiency of compost and biochar amendments

The aim of this study was to test the impact of compost and biochar, with or without earthworms, on the mobility and availability of metals, and on the growth of grass to re-vegetate contaminated soil from the Parys Mountain mining site, Anglesey. We also determined if the addition of earthworms compromises remediation efforts.In a laboratory experiment, contaminated soil (1343 mg Cu kg^?1, 2511 mg Pb kg^?1 and 262 mg Zn kg^?1) was remediated with compost and/or biochar. After 77 days Lumbricus terrestris L. earthworms were added to the treatment remediated with both compost and biochar, and left for 28 days. L. terrestris was not able to survive in the biochar, compost or unamended treatments. A germination and growth bioassay, using Agrostis capillaris (Common Bent) was then run on all treatments for 28 days.The combination of biochar and compost decreased water soluble Cu (from 5.6 to 0.2 mg kg^?1), Pb (from 0.17 to less than 0.007 mg kg^?1) and Zn (from 3.3 to 0.05 mg kg^?1) in the contaminated soil and increased the pH from 2.7 to 6.6. The addition of L. terrestris to this treatment had no effect on the concentration of the water soluble metals in the remediated soil.The compost was the only treatment that resulted in germination and growth of A. capillaris suitable for re-vegetation purposes. However, the combination of compost and biochar (with or without L. terrestris) produced the lowest concentrations of Cu (8 mg kg^?1) and Zn (36 mg kg^?1) in the aboveground biomass, lower than the compost treatment (15 mg Cu kg^?1 and 126 mg Zn kg^?1).The addition of biochar and compost both separately and as co-amendments was effective in reducing the mobility and availability of metals. The addition of L. terrestris did not re-mobilise previously sequestered metals.     

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

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

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.     

The effects of fresh and stabilized pruning wastes on the biomass,structure and activity of the soil microbial community in a semiarid climate

The incorporation of organic amendments from pruning waste into soil may help to mitigate soil degradation and to improve soil fertility in semiarid ecosystems. However, the effects of pruning wastes on the biomass, structure and activity of the soil microbial community are not fully known. In this study, we evaluate the response of the microbial community of a semiarid soil to fresh and composted vegetal wastes that were added as organic amendments at different doses (150 and 300 t ha⁻¹) five years ago. The effects on the soil microbial community were evaluated through a suite of different chemical, microbiological and biochemical indicators, including enzyme activities, community-level physiological profiles (CLPPs) and phospholipid fatty acid analysis (PLFA). Our results evidenced a long-term legacy of the added materials in terms of soil microbial biomass and enzyme activity. For instance, cellulase activity reached 633 μg and 283 μg glucose g⁻¹ h⁻¹ in the soils amended with fresh and composted waste, respectively. Similarly, bacterial biomass reached 116 nmol g⁻¹ in the soil treated with a high dose of fresh waste, while it reached just 66 nmol g⁻¹ in the soil amended with a high dose of composted waste. Organic amendments produced a long-term increase in microbiological activity and a change in the structure of the microbial community, which was largely dependent on the stabilization level of the pruning waste but not on the applied dose. Ultimately, the addition of fresh pruning waste was more effective than the application of composted waste for improving the microbiological soil quality in semiarid soils.     

Evaluation of an optimal extraction method for measuring d-ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) in agricultural soils and its association with soil microbial CO2 assimilation

Assimilating atmospheric carbon (C) into terrestrial ecosystems is recognized as a primary measure to mitigate global warming. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the dominant enzyme by which terrestrial autotrophic bacteria and plants fix CO₂. To investigate the possibility of using RubisCO activity as an indicator of microbial CO₂ fixation potential, a valid and efficient method for extracting soil proteins is needed. We examined three methods commonly used for total soil protein extraction. A simple sonication method for extracting soil protein was more efficient than bead beating or freeze–thaw methods. Total soil protein, RubisCO activity, and microbial fixation of CO₂ in different agricultural soils were quantified in an incubation experiment using ¹⁴C-CO₂ as a tracer. The soil samples showed significant differences in protein content and RubisCO activity, defined as nmol CO₂ fixed g⁻¹ soil min⁻¹. RubisCO activities ranged from 10.68 to 68.07 nmol CO₂ kg⁻¹ soil min⁻¹, which were closely related to the abundance of cbbL genes (r = 0.900, P = 0.0140) and the rates of microbial CO₂ assimilation (r = 0.949, P = 0.0038). This suggests that RubisCO activity can be used as an indicator of soil microbial assimilation of atmospheric CO₂.     

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.