土壤酶对重金属污染的响应及指示研究进展

土壤酶在关键元素生物地球化学循环、动植物健康维持、环境污染净化等方面起着不可替代的重要作用,同时还是土壤污染程度评价的辅助指标之一。然而,由于土壤物理、化学和生物学性质的差异,以及研究方法的多样化,导致重金属与土壤酶活性之间的关系十分复杂,阻碍了土壤酶在土壤质量和健康评价中的应用。系统阐述了重金属污染对土壤酶的生态毒理效应,及其对土壤酶催化动力学特征的影响,构建了土壤-重金属-微生物对土壤酶作用的概念模型,并探讨未来的研究趋势和方向。土壤酶活性测定高效、便宜,且对重金属污染敏感,是极具潜力的土壤重金属污染评价的生物学指标,但仅采用土壤酶活性可能高估或低估重金属的生态毒性,加之当前对土壤酶的选择、活性的测定均缺乏统一的标准,致使难以建立重金属毒性阈值与土壤理化性质或土壤重金属有效性之间的定量关系,最终导致土壤酶在土壤污染生态风险评价中存在争议。未来亟需通过新技术和数学模型,深入揭示不同类型土壤中酶对重金属胁迫的响应机理,构建土壤性质与毒性阈值关系的经验模型,可为加强土壤酶在土壤质量和健康评价中的应用提供重要的理论依据。     

Structure and function of the soil microbial community in microhabitats of a heavy metal polluted soil

 Particle-size fractionation of a heavy metal polluted soil was performed to study the influence of environmental pollution on microbial community structure, microbial biomass, microbial residues and enzyme activities in microhabitats of a Calcaric Phaeocem. In 1987, the soil was experimentally contaminated with four heavy metal loads: (1) uncontaminated controls; (2) light (300 ppm Zn, 100 ppm Cu, 50 ppm Ni, 50 ppm V and 3 ppm Cd); (3) medium; and (4) heavy pollution (two- and threefold the light load, respectively). After 10 years of exposure, the highest concentrations of microbial ninhydrin-reactive nitrogen were found in the clay (2–0.1 μm) and silt fractions (63–2 μm), and the lowest were found in the coarse sand fraction (2,000–250 μm). The phospholipid fatty acid analyses (PLFA) and denaturing gradient gel electrophoresis (DGGE) separation of 16S rRNA gene fragments revealed that the microbial biomass within the clay fraction was predominantly due to soil bacteria. In contrast, a high percentage of fungal-derived PLFA 18 : 2ω6 was found in the coarse sand fraction. Bacterial residues such as muramic acid accumulated in the finer fractions in relation to fungal residues. The fractions also differed with respect to substrate utilization: Urease was located mainly in the <2 μm fraction, alkaline phosphatase and arylsulfatase in the 2–63 μm fraction, and xylanase activity was equally distributed in all fractions. Heavy metal pollution significantly decreased the concentration of ninhydrin-reactive nitrogen of soil microorganisms in the silt and clay fraction and thus in the bulk soil. Soil enzyme activity was reduced significantly in all fractions subjected to heavy metal pollution in the order arylsulfatase >phosphatase >urease >xylanase. Heavy metal pollution did not markedly change the similarity pattern of the DGGE profiles and amino sugar concentrations. Therefore, microbial biomass and enzyme activities seem to be more sensitive than 16S rRNA gene fragments and microbial amino-sugar-N to heavy metal treatment. Received: 21 January 2000     

藏中矿区重金属污染对土壤微生物学特性的影响

土壤微生物对土壤重金属污染反应敏感,是探讨矿区土壤重金属污染生态效应的有效指标之一。通过野外调查与采样和室内分析,研究了藏中矿区重金属污染对土壤蔗糖酶、脲酶、脱氢酶和酸性磷酸酶活性、微生物生物量C（MBC）、N（MBN）和P（MBP）、土壤基础呼吸、代谢商（qCO2）及可矿化N的影响。研究表明,矿区土壤重金属Cu、Zn、Pb、Cd全量和有效含量均高于对照土壤;随着矿区土壤重金属含量增加,土壤酶活性、微生物量C、N和P、可矿化N均逐渐降低,土壤基础呼吸和qCO2则逐渐升高;土壤重金属与土壤蔗糖酶活性、脲酶活性、脱氢酶活性、酸性磷酸酶活性、MBC、MBN、土壤基础呼吸、qCO2及可矿化N具有显著的线性相关;脱氢酶活性对土壤重金属污染最为敏感,表明脱氢酶活性可作为藏中矿区土壤环境质量变化的有效指标。     

Microbial biomass,activity, and organic matter accumulation in soils contaminated with heavy metals

Chemical characteristics and some parameters related to biological components were determined in 16 soils from a fairly homogeneous area in the north of Italy, contaminated with different levels of heavy metals. Correlation analysis of the parameters studied showed close positive relationships among the metals and with the organic C content in the soils studied. Negative relationships were observed among the heavy metals, soil respiration, and the ratio between evolved CO₂–C and microbial biomass C per unit time (specific respiratory activity). This was ascribed to an adverse heavy metal effect on the soil microflora, which appeared to increase the accumulation of organic matter as the heavy metal content increased, probably because the biomass was less effective in mineralising soil organic matter under these conditions.     

Microbiological rates and enzyme activities as indicators of functionality in soils affected by the Aznalcóllar toxic spill

This study focused on the potential of using soil enzyme activities and general microbiological rates (respiration, N-mineralisation, nitrification) to evaluate the quality of soils affected by a pyrite mud spill which contained high concentrations of heavy metals. The quality of soils after restoration was estimated by comparing enzyme activities and general microbiological rates in three different types of experimental field plots: (i) non-polluted, (ii) polluted but restored, and (iii) polluted but un-restored soils. Non-polluted soils showed the highest levels of enzyme activity. Significant differences were detected for acid phosphatase, β-glucosidase and urease activities between all types of plots. However, arylsulfatase and alkaline phosphatase activities showed no significant differences between the restored plots and polluted but un-restored plots. Geometric mean statistics were used as an index of soil quality in terms of overall: (i) bioavailable heavy metal concentrations, (ii) assayed enzyme activities, and (iii) general microbiological rates, in order to compare plots differing in the degree of pyritic mud pollution. The results indicate that it is important to consider these three criteria in to estimate the soil quality of heavy-metal contaminated soils. Typically, enzyme activities were negatively correlated with bioavailable Cd, Cu and Zn concentrations, but positively with soil pH values. In contrast, pH values were negatively correlated with bioavailable concentrations of Cd, Cu and Zn. It is unclear if the generalised lower enzyme activities found in restored soils, compared to non-polluted soils, is promoted by pH or bioavailable heavy metals concentrations, or a combination of both.     

重金属污染对水稻土微生物及酶活性影响研究进展

水稻土受到重金属污染不仅影响水稻的产量品质，而且对水稻土微生物及酶活性的影响不容忽视。本文系统综述了水稻土重金属污染的来源，重金属污染对水稻土微生物生物量、种群数量、群落结构以及土壤酶活性的影响，并针对重金属污染对水稻土微生态效应研究的不足提出了未来应该研究的重点和方向，指出：①加强水稻-重金属-微生物三者相互作用、相互影响方面的研究；②在研究重金属污染与水稻土土壤微生物生态特征的关系的基础上，加强对重金属、土壤理化性状和水稻等因素进行综合并定量化分析，将是明确重金属对土壤微生物生态特性的影响及相关机理的关键；③应用分子生物学方法以及系统生物学方法，促进重金属污染胁迫下水稻土微生物活性及功能的演变规律及响应适应过程；④加强基于长期定位实验的研究，在较长的时间尺度和较大的空间尺度上认识水稻土生态系统在重金属胁迫下的演变规律和机制；⑤重金属污染对水稻土酶活性的研究应重点从机理方面入手，注重结合土壤酶的动力学参数和热力学参数，深化土壤酶与复合污染的作用机理，进一步揭示复合污染致毒途径及其机理，同时借助分子手段，探索重金属污染水稻土中更多未发现的酶的特性，寻找更加敏感、更能普遍推广的重金属污染土壤的综合性指标，以期为重金属污染水稻土的风险评价和生物修复提供科学依据。     

铅锌矿区污染土壤微生物活性研究

通过野外调查和采样分析，研究了浙江衢州铅锌矿区土壤的微生物、土壤酶活性及植物重金属积累特性。结果表明：矿区污染区土壤Ph、Zn、Cd、Cu全量的平均值分别是对照土壤的267．8倍、132．6倍、41．8倍、17．0倍。矿区植物体内重金属含量与土壤重金属全量和有效态含量呈显著正相关。矿区土壤随着重金属含量的增加，土壤微生物生物量碳逐渐降低，而土壤基础呼吸、微生物代谢商则升高，矿区中心污染土壤微生物生物量碳只有对照土壤的72％，而基础呼吸和微生物代谢商分别是对照土壤的1．6倍和2．3倍。铅锌矿口附近污染区土壤酶活性较低，对照土壤的各种酶活性最高。其中土壤脱氧酶的活性变化最大，作为矿区重金属污染的指标更灵敏。     

铅锌银尾矿区土壤微生物活性及其群落功能多样性研究

通过对浙江省天台铅锌银尾矿区土壤微生物活性指标以及微生物群落功能多样性研究 ,结果表明 ,尾矿污染区土壤几种重金属含量比非矿区土壤有明显的增加。尾矿区土壤微生物特征发生了显著的变化 ,微生物生物量和可培养细菌数量显著降低 ,但土壤基础呼吸和微生物代谢商 (qCO2 )值却明显升高。Bi olog测试结果显示 ,随着重金属污染程度的加剧其土壤微生物群落结构发生了相应变化 ,尾矿区土壤微生物群落代谢剖面 (AWCD)及群落丰富度、多样性指数均显著低于非矿区土壤 ,且供试土壤间均达极显著水平差异 (p <0 .0 1) ,表明尾矿区重金属污染引起了土壤微生物群落功能多样性的下降 ,减少了能利用有关碳源底物的微生物数量、降低了微生物对单一碳源底物的利用能力     

重金属复合污染下红壤微生物活性及其群落结构的变化

在室内培养条件下,应用4因素5水平二次正交回归旋转组合设计方案,对重金属复合污染红壤的微生物活性及群落功能多样性进行了研究。结果表明,在Cu、Zn、Pb、Cd复合污染生境中重金属的微生物毒性效应发挥主要由Cd、Cu两元素决定,其生物毒性顺序表现为:Cd>Cu>Zn>Ph,红砂泥中表现较为突出。与单一镉、单一铅污染处理相比,重金属复合污染对供试红壤微生物生物活性及其群落功能多样性的影响并非仅仅表现出简单的加和作用,同时还存在协同作用和拮抗作用。典型相关分析显示,红壤微生物活性与重金属含量之间关系密切,第一对典型变量的相关系数r达0.885 8、0.932 8,均达极显著水平(p<0.001), 说明重金属复合污染与红壤微生物间存在较为稳定的数量反馈机制,红壤总体微生物活性指标能较好地反映供试红壤重金属复合污染状况,可作为重金属污染红壤环境质量评价及量化分类的有效指标。     

南方几种水稻土重金属污染下的土壤呼吸及微生物学效应

采集南方几种重金属污染下的水稻土,通过室内培养的方法研究土壤CO2排放的动态变化以及微生物学指标的差异。结果表明,在60d的培养期内,前7d土壤呼吸速率较高,占了整个排放量的30．89％～64．37％,并且这一阶段重金属对土壤呼吸速率的影响最大。重金属对土壤微生物生物量的影响表现出增加、抑制与无显著性差异的结果,而重金属对微生物熵及微生物代谢熵（qCO2）的影响却是极显著的,同时表现出增加与降低的不同结果。这说明土壤呼吸以及不同的微生物学指标,在长期的复合重金属污染条件下,其表现并不一致,微生物熵与代谢熵用于基本性质差异较大的土壤时,对重金属的响应更为灵敏。此外,土壤重金属的累积还能提高土壤中有机碳的含量。     

重金属Cd、Zn、Cu和Pb复合污染对土壤生物活性的影响

通过野外土样采集及室内培养试验(25℃),研究了云南东川铜矿区土壤酶和微生物特征,以及模拟重金属Cd、Zn、Cu、Pb复合污染对土壤微生物和酶活性的影响。结果表明,矿区土壤(距矿口0~800 m)重金属污染严重,Pb、Cd、Zn、Cu全量和有效含量是对照土壤(距矿口10 000 m)的3.7~141.0倍和2.2~773.2倍;距矿口越近,土壤有机质、有效氮、有效磷和速效钾含量及土壤pH亦越低,土壤酶活性和土壤微生物数量、微生物生物量碳和氮受到的抑制程度也显著增强。与对照土壤相比,距矿口0~800 m的土壤蔗糖酶、脲酶、酸性磷酸酶、过氧化氢酶和脱氢酶活性分别降低25.5%~47.3%、22.6%~74.2%、30.9%~83.1%、16.7%~69.1%和34.6%~92.3%;细菌、放线菌和真菌数量分别较对照下降30.5%~80.1%、8.1%~49.9%和3.3%~8.3%。土壤酶中的酸性磷酸酶和过氧化氢酶,土壤微生物中的细菌对重金属污染较为敏感。恒温(25℃)培养试验中,低量的Cd、Zn、Cu、Pb复合污染刺激了土壤酶活性和细菌、真菌、放线菌、微生物生物量碳和氮的数量,但高量的Cu、Zn、Pb、Cd复合污染使土壤酶活性、细菌、真菌、放线菌、微生物生物量碳和氮均显著下降。重金属Cd、Zn、Cu、Pb之间存在着一定的协同或拮抗作用,Cd、Zn、Cu和Pb之间在微生物生物量碳和氮上表现出明显的协同效应,Pb与Cd、Zn、Cu对细菌数量的复合效应机制为拮抗效应,Cd、Zn、Cu和Pb对真菌数量和放线菌数量的复合效应机制表现为协同效应和拮抗效应并存。     

粤北铅锌矿区土壤生态系统微生物特征及其重金属含量

粤北大宝山金属矿产资源的开发给生态环境带来了严重危害。对该铅锌矿区土壤的微生物特征进行研究。结果表明:五节芒植物中的元素含量表现为Zn>Pb>Cu>Cd,Zn与土壤元素的相关性最为显著,其次为Pb。与对照土壤相比,矿区土壤的微生物基础呼吸作用增强,但微生物生物量却显著降低,微生物生理生态参数Cmic/Corg、qCO2值明显升高。随着矿区土壤的重金属含量增加,矿区(1#、2#)土壤的生化作用强度明显下降,生化作用表现为与土壤重金属含量呈显著负相关。与非矿区(6#)土壤相比,其中土壤氨化作用、硝化作用、固N作用和纤维素分解作用强度分别下降43.19%～70.01%,70.71%～92.02%,58.54%～87.76%和55.00%～79.60%。生化作用表现为与土壤中重金属含量呈显著负相关。土壤微生物活性下降是矿区土壤生态系统遭受破坏的重要标志之一,也是矿区土壤微生物生态演变的重要因素之一。土壤微生物活性降低削弱了矿区土壤中C、N营养元素的循环速率和能量流动。     

Suitability of soil microbial parameters as indicators of heavy metal pollution

Several microbial parameters (microbial biomass, respiration, dehydrogenase, phosphatase, sulphatase, glucosidase, protease and urease activities) were measured in soils from five sites located in urban green areas close to roads differing in traffic density. Our aims were to evaluate the suitability of such parameters as field biomarkers of stress induced by heavy metal pollution, and to compare results obtained by single microbial parameters with results given by an index expressing the average microbial (AME) response of the microbial community. Data showed that all parameters were significantly reduced in the sites characterized by the highest load of metals in soil. Dehydrogenase, sulphatase, glucosidase activities and respiration, declined exponentially with increasing metal concentration, whereas phosphatase activity and AME decreased following a sigmoidal type relationship. In contrast, protease, urease and microbial biomass were not significantly correlated with soil metal concentration. Microbial parameters differed both in sensitivity to critical metal concentrations and in the rate of decline at increasing metal loads in soil. Due to the complex interplay of chemical, physical and biological factors which influence microbial activities and biomass, the proposed index (AME) appeared more suitable than single microbial parameters for a biomonitoring study of this type.     

长期重金属胁迫下的土壤微生物特征: 沈阳张士污灌区的案例研究

Soil samples were collected from Zhangshi Wastewater Irrigation Area in the suburb of Shenyang City, China, an area with a 30-year irrigation history with heavy metal-containing wastewater. The chemical properties and microbial characteristics of the soils were examined to evaluate the present situation of heavy metal pollution and to assess the soil microbial characteristics under long-term heavy metal stress. In light of the National Environmental Quality Standards of China, the soil in the test area was heavily polluted by Cd and to a lesser degree by Zn and Cu, even though wastewater irrigation ceased in 1993. Soil metabolic quotient （qCO2） had a significant positive correlation, while soil microbial quotient （qM） had a negative correlation with content of soil heavy metals. Soil microbial biomass carbon （MBC） had significantly negative correlation with Cd, but soil substrate~induced respiration （SIR）, dehydrogenase activity （DHA）, cellulase activity, and culturable microbial populations had no persistent correlations with soil heavy metal content. Soil nutrients, except for phosphorous, showed positive effects on soil microbial characteristics, which to a certain degree obscured the adverse effects of soil heavy metals. Soil Cd contributed more to the soil microbial characteristics, but qM and qCO2 were more sensitive and showed persistent responses to heavy metals stress. It could be concluded that qM and qCO2 can be used as bioindicators of heavy metal pollution in soils.     

Temporal response of soil biochemical properties in a pastoral soil after cultivation following high application rates of undigested sewage sludge

Soil biochemical properties were measured annually between 1995 and 1999 in soil from an 8-ha site that had received over 1,000 wet tonnes ha^–1 undigested sewage sludge, 1–4 years earlier. Basal respiration generally declined with time and was usually greatest in the untreated control area. This trend was attributed to a similar trend in soil moisture content. In contrast, microbial biomass C increased with time and also generally increased with sludge treatment age. Microbial biomass C, and to a lesser extent sulphatase activity, accurately predicted the order of sludge application to the site. This was perceived as a function of time since tillage and pasture establishment, with activities increasing in parallel to the build up of C residues in the soil, and not an effect of sludge or its composition. Except immediately after sludge application, there was no effect on N mineralisation and nitrification. None of the biochemical properties was strongly correlated with heavy metal concentrations. Our results suggest that there was little effect on soil biochemical properties, either adverse or beneficial, of adding raw sewage sludge to this site. Although a companion study showed considerable mobility and plant uptake of heavy metals, this difference could mainly be attributed to a different sampling strategy and the effects of intensive liming of the site.     

Snow removal alters soil microbial biomass and enzyme activity in a Tibetan alpine forest

Projected future decreases in snow cover associated with global warming in alpine ecosystems could affect soil biochemical cycling. To address the objectives how an altered snow removal could affect soil microbial biomass and enzyme activity related to soil carbon and nitrogen cycling and pools, plastic film coverage and returning of melt snow water were applied to simulate the absence of snow cover in a Tibetan alpine forest of western China. Soil temperature and moisture, nutrient availability, microbial biomass and enzyme activity were measured at different periods (before snow cover, early snow cover, deep snow cover, snow cover melting and early growing season) over the entire 2009/2010 winter. Snow removal increased the daily variation of soil temperature, frequency of freeze–thaw cycle, soil frost depth, and advanced the dates of soil freezing and melting, and the peak release of inorganic N. Snow removal significantly decreased soil gravimetric water, ammonium and inorganic N, and activity of soil invertase and urease, but increased soil nitrate, dissolve organic C (DOC) and N (DON), and soil microbial biomass C (MBC) and N (MBN). Our results suggest that a decreased snow cover associated with global warming may advance the timing of soil freezing and thawing as well as the peak of releases of nutrients, leading to an enhanced nutrient leaching before plant become active. These results demonstrate that an absence of snow cover under global warming scenarios will alter soil microbial activities and hence element biogeochemical cycling in alpine forest ecosystems.     

Changes in Microbial Functional Diversity and Activity in Paddy Soils Irrigated with Industrial Wastewaters in Bandung, West Java Province, Indonesia

Characteristics, such as microbial biomass, basal respiration, and functional diversity of the microbial communities, were investigated in paddy soils located in Bandung, West Java Province, Indonesia, that have been heavily polluted by industrial effluents for 31 years. Paddy soil samples (10?C20 cm) were taken from two sites: polluted soils and unpolluted soils (as control sites). The polluted soils contained higher salinity, higher sodicity, higher nutrient contents, and elevated levels of heavy metals (Cr, Mn, Ni, Cu, and Zn) than unpolluted soils. Soil physicochemical properties, such as maximum water holding capacity, exchangeable sodium percentage, sodium adsorption ratio, and swelling factor, in polluted soils were much greater than those in unpolluted soils (P?<?0.05). Changes in the physical and chemical soil properties were reflected by changes in the microbial communities and their activities. BIOLOG analysis indicated that the functional diversity of the microbial community of polluted soils increased and differed from that of unpolluted soils. Likewise, the average rate of color development (average well color development), microbial biomass (measured as DNA concentration), and the soil CO₂ respiration were higher in polluted soils. These results indicate that major changes in the chemical and physical properties of paddy soils following the application of industrial wastewater effluents have had lasting impacts on the microbial communities of these soils. Thus, the increased activity, biomass, and functional diversity of the microbial communities in polluted soils with elevated salinity, sodicity, and heavy metal contents may be a key factor in enhancing the bioremediation process of these heavily polluted paddy soils.     

采煤沉陷复垦区重金属污染与土壤酶活性的关系

以淮南新庄孜矿采煤沉陷复垦区为研究区域,对脲酶、磷酸酶、蔗糖酶、蛋白酶、纤维素酶、过氧化氢酶、多酚氧化酶7种土壤酶进行了测试分析。结果发现,该区域土壤酶活性受到不同程度的抑制,且7种土壤酶对重金属的敏感性不一致。采用因子分析技术提取主成分,建立了复合污染土壤总体酶活性评价指标,评价了该地区土壤重金属污染程度,并与通过灰色聚类法分析得到的土壤重金属污染等级进行了比对分析,研究了采煤沉陷区土壤酶活性与重金属污染之间的赋存关系,进一步证明在采煤沉陷复垦区这一特殊土地利用条件下,采用土壤酶活性综合指标评价土壤重金属污染是可行的。研究表明,土壤酶总体活性评价指标作为一种简单易行的评价手段,为新庄孜采煤沉陷复垦区及相似矿区复垦地的重金属污染修复提供参考和理论依据。     

Effect of arsenic contamination on bacterial and fungal biomass and enzyme activities in tropical arsenic-contaminated soils

The importance of assessing the impacts of soil arsenic (As) contamination on microbial properties lay on the fact that microbes are instrumental in nutrient cycling and are therefore indicators of soil quality. In this study, soil chemical extraction methods were used to extract labile and freely exchangeable As (water-soluble As and sodium bicarbonate-extractable As), amorphous/crystalline Fe and Mn oxide-bound As (acid ammonium oxalate-extractable As and hydroxylamine hydrochloride-extractable As), and their impacts on microbial biomass (microbial biomass C, total bacterial and fungal biomass, active bacterial and fungal biomass), enzyme activities representing four major soil biogeochemical cycles, i.e., C (β-glucosidase activity), N (urease activity), P (acid phosphomonoesterase activity), S (acryl-sulfatase activity), and microbial activity (fluorescein diacetate hydrolysis and dehydrogenase activity) were investigated in As-contaminated soils of Ambagarh Chauki block, Chhattisgarh, Central India. The results revealed that the majority of the As in soils resided in the Fe/Mn oxide-bound fraction. The microbial biomass C, total and active fungal biomass, and enzyme activities were significantly inhibited by all the forms of As. However, water-soluble As, even though occupying only a small portion of the total As (0.9–2.9 %), exerted the greatest impact. Interestingly, total and active bacterial biomass was not significantly affected by As toxicity, suggesting their resistance to As. Urease activity was not affected by As pollution.     

Soil moisture pre-treatment effects on enzyme activities as indicators of heavy metal-contaminated and reclaimed soils

Heavy metal contamination can inhibit soil functions but it is often difficult to determine the degree of pollution or when soil reclamation is complete. Enzyme assays offer potential as indicators of biological functioning of soils. However, antecedent water content of soil samples may affect the outcome of biological measurements. In Mediterranean regions, for much of the year ‘field moist’ surface soil can have water content similar to that of air-dry samples. The objectives of this study were to: (1) determine the sensitivity of a range of enzyme assays to detect the degree of pollution from a heavy metal mine spill; (2) evaluate rewetting field-dry soil as a pre-treatment for enzyme assays; and (3) test multivariate analysis for improving discrimination between polluted, reclaimed and non-polluted soils. The Aznalcóllar mining effluent spill provided a unique opportunity to address these objectives. This accident released toxic, heavy metal-contaminated (As, Bi, Cd, Cu, Pb, Tl, Zn…) and acid tailings into the Guadiamar watershed (SW Spain) in 1998, severely affecting the riparian zone along more than 4000 ha. Contaminated soils were collected from the highly polluted upper watershed and less polluted lower watershed along with reclaimed soil at both sites. Enzyme activities (phosphatases, arylsulfatase, β-glucosidase, urease and dehydrogenase) were assessed on both field-moist samples and soils rewetted to 80% of water-holding capacity and then incubated at 21 °C for 7 d prior to the assay. The reclaimed soils had higher activities than polluted soils but, typically, 1.5-3 times lower levels of activity than the non-polluted soil. Regardless of the moisture pre-treatment, all enzymes showed significant effects due to pollution, with urease and β-glucosidase showing the greatest discrimination between degrees of contamination. In general, rewetting field-dried soils increased activities on non-polluted and reclaimed soils which improved discrimination with polluted soils. Another method to increase the potential of soil enzyme activities to detect soil contamination could be to combine them in multivariate analysis, which provides a more holistic representation of the biochemical and microbial functionality of a soil.