Microbial activity and hydrolase synthesis in long-term Cd-contaminated soils

Alkaline and acid phosphomonoesterase, β-glucosidase, arylsulfatase, protease and urease activities, CO₂-C evolution and ATP content were monitored in long-term Cd-contaminated (0-40 mg Cd kg⁻¹ dry weight soil) sandy soils, kept under maize or ‘set aside’ regimes, amended with plant residues. The organic matter input increased soil respiration, ATP contents and hydrolase activities in all soils. However, the Cd-contaminated soils had significantly higher metabolic quotients (qCO₂), as calculated by the CO₂-to-ATP ratio, and significantly lower hydrolase activities and hydrolase activity-to-ATP ratios for alkaline phosphomonoesterase, arylsulfatase and protease activities, compared with the respective uncontaminated soils. The ratios between acid phosphomonoesterase, β-glucosidase and urease activities and ATP were unaffected. A significantly higher qCO₂/μ ratio, an expression of maintenance energy, was observed in most of the contaminated soils, indicating that more energy was required for microbial synthesis in the presence of high Cd concentrations. It was concluded that exposure to high Cd concentrations led to a less efficient metabolism, which was responsible for lower enzyme activity and synthesis and lower hydrolase activity-to-ATP ratios observed in these Cd-contaminated soils.     

Hydrolase activity, microbial biomass and community structure in long-term Cd-contaminated soils

Long-term effects of high Cd concentrations on enzyme activities, microbial biomass and respiration and bacterial community structure of soils were assessed in sandy soils where Cd was added between 1988 and 1990 as Cd(NO₃)₂ to reach concentrations ranging from 0 to 0.36 mmol Cd kg⁻¹ dry weight soil. Soils were mantained under maize and grass cultivation, or ‘set-aside’ regimes, for 1 year. Solubility of Cd and its bioavailability were measured by chemical extractions or by the BIOMET bacterial biosensor system. Cadmium solubility was very low, and Cd bioavailability was barely detectable even in soils polluted with 0.36 mmol Cd kg⁻¹. Soil microbial biomass carbon (B_C) was slightly decreased and respiration was increased significantly even at the lower Cd concentration and as a consequence the metabolic quotient (qCO₂) was increased, indicating a stressful condition for soil microflora. However, Cd-contaminated soils also had a lower total organic C (TOC) content and thus the microbial biomass C-to-TOC ratio was unaffected by Cd. Alkaline phosphomonoesterase, arylsulphatase and protease activities were significantly reduced in all Cd-contaminated soils whereas acid phosphomonoesterase, β-glucosidase and urease activites were unaffected by Cd. Neither changes in physiological groups of bacteria, nor of Cd resistant bacteria could be detected in numbers of the culturable bacterial community. Denaturing gradient gel electrophoresis analysis of the bacterial community showed slight changes in maize cropped soils containing 0.18 and 0.36 mmol Cd kg⁻¹ soil as compared to the control. It was concluded that high Cd concentrations induced mainly physiological adaptations rather than selection for metal-resistant culturable soil microflora, regardless of Cd concentration, and that some biochemical parameters were more sensitive to stress than others.     

Short-term effects of dairy slurry amendment on carbon sequestration and enzyme activities in a temperate grassland

Land application of animal wastes from intensive grassland farming has resulted in growing environmental problems relating to greenhouse gas emissions, ammonia volatilisation, and nitrate and phosphorus leaching into surface and groundwater. We examined the short-term effects of dairy slurry amendment on carbon sequestration and enzyme activities in a temperate grassland (Southwest England). Slurry was collected from cows fed either on perennial ryegrass (C₃) or maize (C₄) silages. Fifty m³ ha⁻¹ of each of the obtained C₃ or C₄ slurries (δ¹³C=−30.7 and −21.3‰, respectively) were applied to a C₃ pasture soil with δ¹³C of −30.0±0.2‰. We found that water soluble organic carbon (WSOC) content was two to three times higher in the slurry amended plots compared with the unamended control. No significant change in the soil microbial biomass (SMB) carbon content was observed in the four weeks (772 h) following slurry application. Natural abundance ¹³C isotope analysis suggested a rapid initial incorporation (>25% within 2 h of application) of slurry-derived C in the SMB-C and WSOC pools of the 0-2 cm layer. Linear relationships were found between slurry-derived C in the whole soil, SMB, and WSOC for the 0-2 cm depth in the soil. Applied slurry-derived C was sequestered in the SMB pool in two phases. The first phase (0-48 h) was dominated by the incorporation of labile slurry C from the liquid phase, whereas beyond 48 h slurry-derived C was mainly from less mobile particulate C. No significant differences between treatments were found for invertase and xylanase. Urease activity was always higher in slurry treatments. Cellobiohydrolase, β-N-acetyl-glucosamidase, β-glucosidase and acid phosphatase activities became significantly higher in slurry treatments after 336 h. However, the observed temporal changes in enzyme activities were not correlated with the amounts of slurry-C incorporated in the SMB and WSOC pool.     

Stability of hydrolytic enzyme activity and microbial phosphorus during storage of tropical rain forest soils

Storage can markedly influence microbial and biochemical properties in soils, yet recommendations for sample storage are based on studies of temperate soils that regularly experience extended cold periods. We assessed the influence of storage conditions on microbial phosphorus and the activity of four hydrolytic enzymes (phosphomonoesterase, phosphodiesterase, β-glucosidase, and N-acetyl-β-d-glucosaminidase) in three lowland tropical forest soils from the Republic of Panama that experience a constant warm temperature. The soils spanned a strong rainfall gradient and contained contrasting physical and chemical properties (pH 3.6-5.9; total carbon 26-50 g C kg⁻¹; clay 33-62%; total phosphorus 0.30-0.60 g P kg⁻¹). Storage treatments were: (i) room temperature (22 °C in the dark), (ii) refrigerated (4 °C in the dark), (iii) air-dried (10 d, 22 °C), and (iv) frozen (−35 °C). There were significant changes in enzyme activities and microbial phosphorus during refrigerated and room temperature storage, although changes were relatively small during the first two weeks. An initial marked decline in enzyme activities for one soil analyzed within 2 h of sampling was attributed to a flush of activity caused by sampling and soil preparation (sieving, etc.). For longer-term storage (>2 weeks), ambient laboratory temperature appeared preferable to freezing and cold storage, because one month of storage caused a marked decline in enzyme activities and microbial phosphorus in one soil. Freezing preserved the activities of some enzymes in some soils at rates comparable to cold or room temperature storage, but caused a marked decline in microbial phosphorus in two soils. Air-drying caused a marked decline in microbial phosphorus and the activity of all enzymes. We therefore conclude that enzyme assays and microbial phosphorus should be determined in tropical forest soils after no more than two weeks storage in the dark at ambient laboratory temperature.     

Exploring the enzymatic landscape: distribution and kinetics of hydrolytic enzymes in soil particle-size fractions

The location of extracellular enzymes within the soil architecture and their association with the various soil components affects their catalytic potential. A soil fractionation study was carried out to investigate: (a) the distribution of a range of hydrolytic enzymes involved in C, N and P transformations, (b) the effect of the location on their respective kinetics, (c) the effect of long-term N fertilizer management on enzyme distribution and kinetic parameters. Soil (silty clay loam) from grassland which had received 0 or 200 kg N ha⁻¹ yr⁻¹ was fractionated, and four particle-size fractions (>200, 200-63, 63-2 and 0.1-2 μm) were obtained by a combination of wet-sieving and centrifugation, after low-energy ultrasonication. All fractions were assayed for four carbohydrases (β-cellobiohydrolase, N-acetyl-β-glucosaminidase, β-glucosidase and β-xylosidase), acid phosphatase and leucine-aminopeptidase using a microplate fluorimetric assay based on MUB-substrates. Enzyme kinetics (V_max and K_m) were estimated in three particle-size fractions and the unfractionated soil. The results showed that not all particle-size fractions were equally enzymatically active and that the distribution of enzymes between fractions depended on the enzyme. Carbohydrases predominated in the coarser fractions while phosphatase and leucine-aminopeptidase were predominant in the clay-size fraction. The Michaelis constant (K_m) varied among fractions, indicating that the association of the same enzyme with different particle-size fractions affected its substrate affinity. The same values of K_m were found in the same fractions from the soil under two contrasting fertilizer management regimes, indicating that the Michaelis constant was unaffected by soil changes caused by N fertilizer management.     

Enzymatic activities in constructed wetlands and di-n-butyl phthalate (DBP) biodegradation

The bioaccumulation of phthalate acid esters (PAEs) from industrial products and their mutagenic action has been suggested to be a potential threat to human health. The effects of the most frequently identified PAE, Di-n-butyl phthalate (DBP), and its biodegradation, were examined by comparison of two small scale plots (SSP) of integrated vertical-flow constructed wetlands. The influent DBP concentration was 9.84 mg l⁻¹ in the treatment plot and the control plot received no DBP. Soil enzymatic activities of dehydrogenase, catalase, protease, phosphatase, urease, cellulase, β-glucosidase, were measured in the two SSP after DBP application for 1 month and 2 months, and 1 month after the final application. Both treatment and control had significantly higher enzyme activity in the surface soil than in the subsurface soil (P<0.001) and greater enzyme activity in the down-flow chamber than in the up-flow chamber (P<0.05). In the constructed wetlands, DBP enhanced the activities of dehydrogenase, catalase, protease, phosphatase and inhibited the activities of urease, cellulase and β-glucosidase. However, urease, cellulase, β-glucosidase activities were restored 1 month following the final DBP addition. Degradation of DBP was greater in the surface soil and was reduced in sterile soil, indicating that this process may be mediated by aerobic microorgansims. DBP degradation fitted a first-order model, and the kinetic equation showed that the rate constant was 0.50 and 0.17 d⁻¹, the half-life was 1.39 and 4.02 d, and the r² was 0.99 and 0.98, in surface and subsurface soil, respectively. These results indicate that constructed wetlands are able to biodegrade organic PAEs such as DBP.     

Differentiating microbial and stabilized β-glucosidase activity relative to soil quality

Previous research has shown that β-glucosidase activity can detect soil management effects and has potential as a soil quality indicator, but mechanisms for this response are not well understood. A significant amount of hydrolytic enzyme activity comes from extracellular (abiontic) activity that is bound and protected by soil colloids. This study was conducted to determine how management affects the kinetics of this enzyme (K_m, substrate affinity, and V_max, maximum reaction velocity) and its degree of stabilization on soil colloids. Soils were sampled from three sites in Oregon, with a paired comparison within each site of a native, unmanaged soil, and a matching soil under agricultural production (>50 years). Microwave radiation (MW) stress was used to denature the β-glucosidase fraction associated with viable microorganisms in these soils as an estimate of abiontic activity. Total activity and V_max were decreased by both management and MW. The results showed that β-glucosidase activity is sensitive to soil management on a variety of soils and environments (135 vs. 190, 80 vs. 111 and 80 vs. 134 μg PNP g⁻¹ h⁻¹ for managed and unmanaged treatments, respectively, at the three study sites in Oregon). The evidence suggests that this sensitivity to management is not (or minimally) due to differences in isoenzymes (K_m generally was unaffected) but rather due to an overall reduction in the amount of enzyme present (V_max decreased) and that this reduction in activity is reflected more from the activity of enzymes in the stabilized fraction than that associated with viable microbial population. Although β-glucosidase activity after MW irradiation appears to be limited as a soil quality indicator, it maybe useful as research tool to separate abiontic from microbial activity ‘biomass’ β-glucosidase activity correlated with microbial biomass C (r=0.42, P<0.05) but MW irradiated, abiontic, activity did not (r=−0.20^NS).     

三江平原岗地白浆土水解酶活性和动力学特性对不同连作农作物的响应

作物种植会对农田生态系统产生一定的影响。大田试验条件下,在黑龙江省853农场岗地白浆土上连续6年种植玉米、大豆、小麦、水稻,研究了土壤理化性质以及土壤中与碳、氮、磷、硫元素转化相关的9种水解酶活性和动力学特性的响应;同时研究了不同作物种植对土壤脲酶、磷酸单酯酶、磷酸二酯酶、芳基硫酸酯酶及β-葡糖苷酶动力学特性的影响。结果表明,大豆连作土壤的有效氮、总碳、总氮、总磷和总硫含量都稍高;大豆处理土壤pH值略低,但其它三种作物种植下的土壤均呈微酸性,差异不显著。土壤水解酶动力学参数对种植作物的反应与表观活性的反应不一致。玉米连作土壤蛋白酶和磷酸单酯酶活性高于其它处理;小麦处理的磷酸二酯酶和芳基硫酸酯酶活性最高,水稻连作土壤蛋白酶、磷酸二酯酶和磷酸三酯酶活性最低。连年种植小麦处理的土壤脲酶、磷酸二酯酶以及芳基硫酸酯酶Vmax显著高于其它处理,小麦连作土壤β-葡萄糖苷酶、脲酶、磷酸二酯酶和芳基硫酸酯酶的Vmax/Km值显著高于其它处理,可以看出在此处理下土壤酶具有较强的催化潜势。     

Attempts to derive EC50 values for heavy metals from land-applied Cu-, Ni-, and Zn-spiked sewage sludge

We established a field trial to assess the impacts on soil biological properties of application of heavy metal-spiked sewage sludge, with the aim of determining toxicity threshold concentrations of heavy metals in soil. Plots were treated with sludges containing increasing concentrations of Cu, Ni and Zn in order to raise the metal concentrations in the soil by 0-200 mg Cu kg⁻¹, 0-60 mg Ni kg⁻¹ and 0-400 mg Zn kg⁻¹, and were then cultivated and sown in ryegrass-clover pasture and monitored annually for 6 years. All biological properties measured (soil basal respiration, microbial biomass C, and sulphatase enzyme activities), except phosphatase activity, increased in all plots over the duration of the experiment. Consequently, it was only possible to assess effects of heavy metals across time if, each year, all data for each metal were normalised by expressing them as percentages of the activities measured in an un-sludged control plot. When this was done, no significant effects of increasing heavy-metal concentrations on basal respiration, microbial biomass C or respiratory quotient (qCO₂) were observed, although total Cu and soil solution Cu were significantly negatively related to microbial biomass C when it was expressed as a proportion of soil total C. None of the properties measured were affected by increasing Ni concentrations. Phosphatase and sulphatase activities were significantly negatively related to increasing Zn concentrations, but not usually to increasing Cu unless they were expressed as a proportion of total C. A sigmoidal dose-response model was used to calculate EC₂₀ and EC₅₀ values using the normalised data, but generally, the model parameters had very large 95% confidence intervals and/or the fits to the model had small R² values. The factors primarily responsible for confounding these results were site and sample variations not accounted for by the normalisation process and the absence of any data points at metal concentrations beyond the calculated EC₅₀ values. In the few instances where reasonable EC₂₀ values could be calculated, they were relatively consistent across properties, e.g., EC₂₀ for total Zn and phosphatase (330 mg kg⁻¹), total Zn and sulphatase (310 mg kg⁻¹), and EC₂₀ for total Cu and sulphatase (140 mg kg⁻¹) and total Cu and microbial biomass C (140 mg kg⁻¹), when both sulphatase and microbial biomass C were expressed as a proportion of total C. Our results suggest that Cu and Zn at the upper concentrations used in this experiment were possibly having adverse effects on some soil biological properties. However, much higher metal concentrations will be needed to accurately calculate EC₂₀ and EC₅₀ and this may not be easily achievable without many applications of sewage sludge, even if the sludge is spiked with heavy metals.     

The influence of time, storage temperature, and substrate age on potential soil enzyme activity in acidic forest soils using MUB-linked substrates and l-DOPA

The purpose of this experiment was to evaluate whether soil storage and processing methods significantly influence measurements of potential in situ enzyme activity in acidic forest soils. More specifically, the objectives were to determine if: (1) duration and temperature of soil storage; (2) duration of soil slurry in buffer; and (3) age of model substrates significantly influence the activity of six commonly measured soil extracellular enzymes using methylumbelliferone (MUB)-linked substrates and l-dihydroxyphenylalanine (l-DOPA). Soil collected and analyzed for enzyme activity within 2 h was considered the best measure of potential in situ enzyme activity and the benchmark for all statistical comparisons. Sub-samples of the same soil were stored at either 4 °C or −20 °C. In addition to the temperature manipulation, soils experienced two more experimental treatments. First, enzyme activity was analyzed 2, 7, 14, and 21 days after collection. Second, MUB-linked substrate was added immediately (i.e. <20 min) or 2 h after mixing soil with buffer. Enzyme activity of soil stored at 4 °C was not significantly different from soil stored at −20 °C. The duration of soil storage was minimal for β-glucosidase, β-xylosidase, and peroxidase activity. N-acetyl-glucosaminidase (NAGase), phosphatase, and phenol oxidase activity appeared to change the most when compared to fresh soils, but the direction of change varied. Likewise, the activities of these enzymes were most sensitive to extended time in buffer. Fluorometric MUB and MUB-linked substrates generally had a 3-day shelf life before they start to significantly suppress reported activities when kept at 4 °C. These findings suggest that the manner in which acidic forest soils are stored and processed are site and enzyme specific and should not initially be trivialized when conducting enzyme assays focusing on NAGase, phosphatase, and phenol oxidase. The activities of β-glucosidase, β-xylosidase, and peroxidase are insensitive to storage and processing methods.     

Enzyme activities in agricultural soils fumigated with methyl bromide alternatives

Pre-plant fumigation of agricultural soils with a combination of methyl bromide (MeBr) and chloropicrin (CP) to control nematodes, soil-borne pathogens and weeds has been a common practice in strawberry (Fragaria X ananassa Duchesne) production since the 1960s. MeBr will be phased out by 2005, but little is known about the impacts of alternative fumigants on soil microbial processes. We investigated the response of microbial biomass and enzyme activities in soils fumigated over two years with MeBr+CP and the alternatives propargyl bromide (PrBr), InLine, Midas and CP. Results were compared to control soils, which were not fumigated for the last 4-5 years for Watsonville and Oxnard, respectively, but had a 10 year history of MeBr+CP fumigation (history soils). Soil samples (0-15 cm) were taken from two sites in the coastal areas of California, USA, in Watsonville and Oxnard, at peak strawberry production after two years of repeated application. In addition to the soil enzymes, the activities of purified reference enzymes of β-glucosidase, acid phosphatase and arylsulfatase were assayed before and after fumigation with MeBr+CP and alternative biocides. At the Oxnard site, microbial respiration significantly decreased in soils fumigated with MeBr+CP (P=0.036), while microbial biomass C and N showed no response to fumigation at both sites. These results may indicate that fumigation promotes the growth of resistant species or that soil microorganisms had recovered at the time of sampling. Repeated soil fumigation with MeBr+CP significantly decreased the activities of β-glucosidase and acid phosphatase at the Watsonville site, and dehydrogenase activity at the Oxnard site. Although, enzyme activities in soils fumigated with PrBr, InLine, Midas and CP were lower compared to the control soil, effects were, in general, not significant. Fumigation with MeBr+CP and alternatives reduced the activities of purified reference enzymes by 13, 76 and 28% for acid phosphatase, β-glucosidase and arylsulfatase, respectively. Mean enzyme protein concentrations in fumigated agricultural soils were 2.93, 0.105, and 2.95 mg protein kg⁻¹ soil for acid phosphatase, β-glucosidase and arylsulfatase, respectively, all lower than in control soils. Organic matter turnover and nutrient cycling, and thus, the long-term productivity of agricultural soils seem unaffected in soils repeatedly fumigated with PrBr, InLine, Midas and CP.     

Shifts in rhizosphere microbial communities and enzyme activity of Poa alpina across an alpine chronosequence

This study quantifies the influence of Poa alpina on the soil microbial community in primary succession of alpine ecosystems, and whether these effects are controlled by the successional stage. Four successional sites representative of four stages of grassland development (initial, 4 years (non-vegetated); pioneer, 20 years; transition, 75 years; mature, 9500 years old) on the Rotmoos glacier foreland, Austria, were sampled. The size, composition and activity of the microbial community in the rhizosphere and bulk soil were characterized using the chloroform-fumigation extraction procedure, phospholipid fatty acid (PLFA) analysis and measurements of the enzymes β-glucosidase, β-xylosidase, N-acetyl-β-glucosaminidase, leucine aminopeptidase, acid phosphatase and sulfatase. The interplay between the host plant and the successional stage was quantified using principal component (PCA) and multidimensional scaling analyses. Correlation analyses were applied to evaluate the relationship between soil factors (C_org, N_t, C/N ratio, pH, ammonium, phosphorus, potassium) and microbial properties in the bulk soil. In the pioneer stage microbial colonization of the rhizosphere of P. alpina was dependent on the reservoir of microbial species in the bulk soil. As a consequence, the rhizosphere and bulk soil were similar in microbial biomass (ninhydrin-reactive nitrogen (NHR-N)), community composition (PLFA), and enzyme activity. In the transition and mature grassland stage, more benign soil conditions stimulated microbial growth (NHR-N, total amount of PLFA, bacterial PLFA, Gram-positive bacteria, Gram-negative bacteria), and microbial diversity (Shannon index H) in the rhizosphere either directly or indirectly through enhanced carbon allocation. In the same period, the rhizosphere microflora shifted from a G⁻ to a more G⁺, and from a fungal to a more bacteria-dominated community. Rhizosphere β-xylosidase, N-acetyl-β-glucosaminidase, and sulfatase activity peaked in the mature grassland soil, whereas rhizosphere leucine aminopeptidase, β-glucosidase, and phosphatase activity were highest in the transition stage, probably because of enhanced carbon and nutrient allocation into the rhizosphere due to better growth conditions. Soil organic matter appeared to be the most important driver of microbial colonization in the bulk soil. The decrease in soil pH and soil C/N ratio mediated the shifts in the soil microbial community composition (bacPLFA, bacPLFA/fungPLFA, G⁻, G⁺/G⁻). The activities of β-glucosidase, β-xylosidase and phosphatase were related to soil ammonium and phosphorus, indicating that higher decomposition rates enhanced the nutrient availability in the bulk soil. We conclude that the major determinants of the microflora vary along the successional gradient: in the pioneer stage the rhizosphere microflora was primarily determined by the harsh soil environment; under more favourable environmental conditions, however, the host plant selected for a specific microbial community that was related to the dynamic interplay between soil properties and carbon supply.     

Enzyme activities in vineyard soils long-term treated with copper-based fungicides

Copper-based fungicides have been applied in vineyard soils for a long time, which has resulted in increasing soil Cu concentration. However, information relating to non-target effects of these fungicides on microorganisms of these soils is scarce. The aim of this study was to determine the potential enzyme activities of vineyard soils in relation to Cu content and evaluate the potential risks of long-term application of Cu-based fungicides. For this purpose, a wide range of soil samples, having different total, exchangeable and bioavailable Cu contents, were collected from six regions of quality wines located in the NW Iberian Peninsula, and the activity of dehydrogenase, β-glucosidase, urease and phosphatase were measured. Overall, the results obtained indicate adverse effects of Cu on dehydrogenase, β-glucosidase and phosphatase activities and an inconsistent effect on urease activity. Threshold Cu concentrations at which changes in the enzyme activities became evident were 150-200 mg total Cu kg⁻¹ and 60-80 mg bioavailable Cu kg⁻¹.     

Intergeneric variability in enzyme production of microfungi from pine litter

Twelve unrelated microfungi from Scots pine (Pinus sylvestris) needle litter were selected to represent supposedly different ecophysiological traits. In order to explore the differences in their involvement in decomposition, activities of their hydrolytic enzymes were compared. Most strains were able to degrade cellulose. The highest exocellulase, β-xylosidase and N-acetylglucosaminidase activities were found in Verticicladium trifidum. Oidiodendron griseum and Allantophomopsis lycopodina were strongly cellulolytic and hemicellulolytic as well. The overlap in the microfungal utilization of cellulose suggests the overlap of nutritional niches of unrelated fungal taxa and competition for carbon sources. Production of the protein-degrading enzymes, arylsulphatase, phosphomonoesterase and phosphodiesterase by litter microfungi was mostly low.     

Enzyme activities and microbial biomass in coastal soils of India

Soil salinity is a serious problem for agriculture in coastal regions, wherein salinity is temporal in nature. We studied the effect of salinity, in summer, monsoon and winter seasons, on microbial biomass carbon (MBC) and enzyme activities (EAs) of the salt-affected soils of the coastal region of the Bay of Bengal, Sundarbans, India. The average pH of soils collected from different sites, during different seasons varied from 4.8 to 7.8. The average organic C (OC) and total N (TN) content of the soils ranged between 5.2-14.1 and 0.6-1.4 g kg⁻¹, respectively. The electrical conductivity of the saturation extract (ECe) of soils, averaged over season, varied from 2.2 to 16.3 dSm⁻¹. The ECe of the soils increased five fold during the summer season (13.8 dSm⁻¹) than the monsoon season (2.7 dSm⁻¹). The major cation and anion detected were Na⁺ and Cl⁻, respectively. Seasonality exerted considerable effects on MBC and soil EAs, with the lowest values recorded during the summer season. The activities of β-glucosidase, urease, acid phosphatase and alkaline phosphatase were similar during the winter and monsoon season. The dehydrogenase activity of soils was higher in monsoon than in winter. Average MBC, dehydrogenase, β-glucosidase, urease, acid phosphatase and alkaline phosphatase activities of the saline soils ranged from 125 to 346 mg kg⁻¹ oven dry soil, 6-9.9 mg triphenyl formazan (TPF) kg⁻¹ oven dry soil h⁻¹, 18-53 mg p-nitro phenol (PNP) kg⁻¹ oven dry soil h⁻¹, 38-86 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 213-584 mg PNP kg⁻¹ oven dry soil h⁻¹ and 176-362 mg PNP g⁻¹ oven dry soil h⁻¹, respectively. The same for the non-saline soils were 274-446 mg kg⁻¹ oven dry soil, 8.8-14.4 mg TPF kg⁻¹ oven dry soil h⁻¹, 41-80 mg PNP kg⁻¹ oven dry soil h⁻¹, 89-134 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 219-287 mg PNP kg⁻¹ oven dry soil h⁻¹ and 407-417 mg PNP kg⁻¹ oven dry soil h⁻¹, respectively. About 48%, 82%, 48%, 63%, 40% and 48% variation in MBC, dehydrogenase activity, β-glucosidase activity, urease activity, acid phosphatase activity and alkaline phosphatase activity, respectively, could be explained by the variation in ECe of saline soils. Suppression of EAs of the coastal soils during summer due to salinity rise is of immense agronomic significance and needs suitable interventions for sustainable crop production.     

Responses of soil chemical and microbial indicators to conservational tillage versus traditional tillage in the North China Plain

This study compared the responses of soil chemical and microbial indicators to the conservational tillage (CT) versus traditional tillage (TT) in a Haplic Cambisol in the North China Plain (NCP). These indicators included soil organic C (SOC), soil total N (STN), soil available P (SAP), cation exchange capacity (CEC), exchangeable Ca²⁺ and Mg²⁺, microbial biomass C (MBC), microbial biomass N (MBN), alkaline phosphomonoesterase (AP), β-glucosidase, N-acetyl-β-glucosaminidase (NAG), nitrate reductase (NR), protease, urease and the geometric mean of the assayed enzymes (GMea). Our results showed that almost all investigated parameters, except the contents of CEC, Ca²⁺, Mg²⁺ and the ratios of GMea/MBN and C/N, were significantly higher under the CT (no-till, NT and reduced-till, RT) than those under the TT, whilst the crop yield was not significantly affected by tillage treatments. Principle component analysis (PCA) showed that the first and second component explained 67.2% and 16.6% of the total variation, respectively. The first component was significantly correlated with GMea, MBC, MBN and β-glucosidase, and effectively discriminated soils under the NT or RT from those under the TT. Our results indicated that the 6-year CT improved the quality of the Haplic Cambisol by enhancing its chemical and microbial properties, whilst GMea, MBC, MBN and β-glucosidase were among the most effective indicators for monitoring these improvements.     

Modelling in situ activities of enzymes as a tool to explain seasonal variation of soil respiration from agro-ecosystems

Understanding in situ enzyme activities could help clarify the fate of soil organic carbon (SOC), one of the largest uncertainties in predicting future climate. Here, we explored the role of soil temperature and moisture on SOM decomposition by using, for the first time, modelled in situ enzyme activities as a proxy to explain seasonal variation in soil respiration. We measured temperature sensitivities (Q₁₀) of three enzymes (β-glucosidase, xylanase and phenoloxidase) and moisture sensitivity of β-glucosidase from agricultural soils in southwest Germany. Significant seasonal variation was found in potential activities of β-glucosidase, xylanase and phenoloxidase and in Q₁₀ for β-glucosidase and phenoloxidase activities but not for xylanase. We measured moisture sensitivity of β-glucosidase activity at four moisture levels (12%–32%), and fitted a saturation function reflecting increasing substrate limitation due to limited substrate diffusion at low water contents. The moisture response function of β-glucosidase activity remained stable throughout the year. Sensitivity of enzymes to temperature and moisture remains one of the greatest uncertainties in C models. We therefore used the response functions to model temperature-based and temperature and moisture-based in situ enzyme activities to characterize seasonal variation in SOC decomposition. We found temperature to be the main factor controlling in situ enzyme activities. To prove the relevance of our modelling approach, we compared the modelled in situ enzyme activities with soil respiration data measured weekly. Temperature-based in situ enzyme activities explained seasonal variability in soil respiration well, with model efficiencies between 0.35 and 0.78. Fitting an exponential response function to in situ soil temperature explained soil respiration to a lesser extent than our enzyme-based approach. Adding soil moisture as a co-factor improved model efficiencies only partly. Our results demonstrate the potential of this new approach to explain seasonal variation of enzyme related processes.     

Shrub encroachment does not reduce the activity of some soil enzymes in Mediterranean semiarid grasslands

Shrub encroachment is a worldwide phenomenon with implications for desertification and global change. We evaluated its effects on the activities of urease, phosphatase and β-glucosidase in Mediterranean semiarid grasslands dominated by Stipa tenacissima by sampling 12 sites with and without resprouting shrubs along a climatic gradient. The presence of shrubs affected the evaluated enzymes at different spatial scales. Soils under S. tenacissima tussocks and in bare ground areas devoid of vascular plants had higher values of phosphatase and urease when the shrubs were present. For the β-glucosidase, this effect was site-specific. At the scale of whole plots (30 m × 30 m), shrubs increased soil enzyme activities between 2% (β-glucosidase) and 22% (urease), albeit these differences were significant only in the later case. Our results indicate that shrub encroachment does not reduce the activity of extracellular soil enzymes in S. tenacissima grasslands.     

Effects of municipal solid waste compost amendments on soil enzyme activities and bacterial genetic diversity

Municipal solid waste (MSW) composts have been used to maintain the long-term productivity of agroecosystems and to protect the soil environment from overcropping, changes in climatic conditions and inadequate management; they also have the additional benefit of reducing waste disposal costs. Since MSW may contain heavy metals and other toxic compounds, amendments cannot only influence soil fertility, but may also affect the composition and activity of soil microorganisms. The effects of MSW compost and mineral N amendments in a 6-year field trial on some physical-chemical properties, enzyme activities and bacterial genetic diversity of cropped plots (Beta vulgaris-Triticum turgidum rotation) and uncropped plots were investigated. The compost was added at the recommended and twice the recommended dosage (12, 24 t ha⁻¹). Amendments of cropped plots with MSW compost increased the contents of organic C from 13.3 to 15.0 g kg⁻¹ soil and total N from 1.55 to 1.65 g kg⁻¹ soil. There were significant increases in dehydrogenase (9.6%), β-glucosidase (13.5%), urease (15.4%), nitrate reductase (21.4%) and phosphatase (9.7%) activities. A significant reduction in protease activity (from 3.6 to 2.8 U g⁻¹ soil) was measured when a double dose of compost was added to the cropped plots. No dosage effect was detected for the other enzymes. Changes in the microbial community, as a consequence of MSW amendment, were minimal as determined using denaturing gradient gel electrophoresis, rDNA internal spacer analysis and amplified ribosomal DNA restriction analysis of bacteria, archaea, actinomycetes, and ammonia oxidizers. This indicates that there was no significant variation in the overall bacterial communities nor in selected taxonomic groups deemed to be essential for soil fertility.     

Logging residue harvest may decrease enzymatic activity of boreal forest soils

Nowadays conventional stem-only harvest where logging residues are left on the site is often displaced by whole-tree harvest, in which logging residues are harvested for use as bioenergy. Logging residues consist of tree branches and tops of stems with needles. The aim of this study was to evaluate the effect of logging residue harvest on soil enzyme activities involved in C, N and P cycling, namely β-glucosidase, β-glucosaminidase, protease and acid phosphatase in relation to other soil characteristics (i.e. soil respiration, net N mineralization, microbial biomass C and N). Soil samples were taken from the humus layer of five study sites, differing in fertility, dominating tree species and time elapsed after treatment. The study sites were Norway spruce (Picea abies, (L.) Karst) and Scots pine (Pinus sylvestris L.) stands in different parts of Finland. Four of the study sites were single-tree experiments, where thinning was performed 4–5 years before this study and 3–4 different doses of logging residues (from 0 up to 37.5 Mg ha⁻¹) were distributed on a circle around a single tree in 3 replicates. The last field experiment had been thinned twice, 23 and 13 years ago; the treatments in 3 replicates were whole-tree harvest and stem-only harvest. In the whole-tree harvest vs. stem-only harvest experiment, activities of β-glucosidase, β-glucosaminidase, acid phosphatase were similar in both treatments. In general, in the single-tree experiment with pine, enzymes raised the activity in response to increasing amount of logging residue. The pattern was less clear for the spruce single-tree experiment, but acid phosphatase and protease activities increased with the increase in amount of logging residue. In general, other soil characteristics were less affected than enzyme activities by logging residue removal; however, in some sites logging residues seemed to increase net C and N mineralization with increasing logging residue amount. Our results suggest that retaining logging residues on the site can increase soil enzyme activities and C and N mineralization.