Interaction of polycyclic aromatic hydrocarbons and heavy metals on soil enzyme

Actions and interactions of heavy metals (cadmium, zinc and plumbum) and polycyclic aromatic hydrocarbons (PAHs) [phenanthrene, fluoranthene, benzo(a)pyrene] on the soil urease and dehydrogenase activity were studied after 49 days exposure. The experimental approach was based on the uniform design which can cut the experiment time and improve the efficiency of experiments. Data treatment was essentially based on the multiple regression technique. The results showed that the action and interaction between heavy metals and PAHs were strongly dependent on the time of pollution. The dehydrogenase exhibits more sensitive to the combined pollution than urease. The negative interaction between Zn and Cd to hydrogenase activity and the combined stimulatory activity of Phenanthrene and Benzo(a)pyrene (or fluoranthene) to soil enzyme were observed. The interactions between Zn (Cd) and phenanthrene towards urease (dehydrogenase) were positive, and the interaction between Zn and benzo(a)pyrene to urease activity was negative. This study corresponds to exploratory phase in order to reveal interaction effects of heavy metals and PAHs on the soil enzyme and then to set up more in-depth analysis to increase progressively the understanding of the ecotoxicological mechanisms involved.     

Zn, Cd and Pb accumulation and arbuscular mycorrhizal colonisation of pennycress Thlaspi praecox Wulf. (Brassicaceae) from the vicinity of a lead mine and smelter in Slovenia

Significant hyperaccumulation of Zn, Cd and Pb in field samples of Thlaspi praecox Wulf. collected from a heavy metal polluted area in Slovenia was found, with maximal shoot concentrations of 14,590 mg kg(-1) Zn, 5960 mg kg(-1) Cd and 3500 mg kg(-1) Pb. Shoot/root ratios of 9.6 for Zn and 5.6 for Cd show that the metals were preferentially transported to the shoots. Shoot bioaccumulation factors exceeded total soil Cd levels 75-fold and total soil Zn levels 20-fold, further supporting the hyperaccumulation of Cd and Zn. Eighty percent of Pb was retained in roots, thus indicating exclusion as a tolerance strategy for Pb. Low level colonisation with arbuscular mycorrhizal fungi (AMF) of a Paris type was observed at the polluted site, whereas at the non-polluted site Arum type colonisation was more common. To our knowledge this is the first report of Cd hyperaccumulation and AMF colonisation in metal hyperaccumulating T. praecox.     

Heavy metal contamination from mining sites in South Morocco: 1. Use of a biotest to assess metal toxicity of tailings and soils

Our work was conducted to investigate the heavy metal toxicity of tailings and soils collected from five metal mines located in the south of Morocco. We used the MetPAD biotest Kit which detects the toxicity specifically due to the heavy metals in environmental samples. This biotest initially developed to assess the toxicity of aquatic samples was adapted to the heterogeneous physico-chemical conditions of anthropogenic soils. Contrasted industrial soils were collected from four abandoned mines (A, B, C and E) and one mine (D) still active. The toxicity test was run concurrently with chemical analyses on the aqueous extracts of tailings materials and soils in order to assess the potential availability of heavy metals. Soil pH was variable, ranging from very acidic (pH 2.6) to alkaline values (pH 8.0-8.8). The tailings from polymetallic mines (B and D) contained very high concentrations of Zn (38,000-108,000 mg kg(-1)), Pb (20,412-30,100 mg kg(-1)), Cu (2,019-8,635 mg kg(-1)) and Cd (148-228 mg kg(-1)). Water-extractable metal concentrations (i.e., soil extracts) were much lower but were highly toxic as shown by the MetPAD test, except for soils from mines A, E and site C3 from mine C. The soil extracts from mine D were the most toxic amongst all the soils tested. On this site, the toxicity of soil water extracts was mainly due to high concentrations of Zn (785-1,753 mg l(-1)), Cu (1.8-82 mg l(-1)) and Cd (2.0-2.7 mg l(-1)). The general trend observed was an increase in metal toxicity measured by the biotest with increasing available metal contents in tailings materials and soils. Therefore, the MetPAD test can be used as a rapid and sensitive predictive tool to assess the heavy metal availability in soils highly contaminated by mining activities.     

Sorption of phenanthrene by soils contaminated with heavy metals

The fate of polycyclic aromatic hydrocarbons (PAHs) in soils with co-contaminants of heavy metals has yet to be elucidated. This study examined sorption of phenanthrene as a representative of PAHs by three soils contaminated with Pb, Zn or Cu. Phenanthrene sorption was clearly higher after the addition of heavy metals. The distribution coefficient (K(d)) and the organic carbon-normalized distribution coefficient (K(oc)) for phenanthrene sorption by soils spiked with Pb, Zn or Cu (0-1000 mg kg(-1)) were approximately 24% larger than those by unspiked ones, and the higher contents of heavy metals added into soils resulted in the larger K(d) and K(oc) values. The enhanced sorption of phenanthrene in the case of heavy metal-contaminated soils could be ascribed to the decreased dissolved organic matter (DOM) in solution and increased soil organic matter (SOM) as a consequence of DOM sorption onto soil solids. Concentrations of DOM in equilibrium solution for phenanthrene sorption were lower in the case of the heavy metal-spiked than unspiked soils. However, the decreased DOM in solution contributed little to the enhanced sorption of phenanthrene in the presence of metals. On the other hand, the sorbed DOM on soil solids after the addition of heavy metals in soils was found to be much more reactive and have far stronger capacity of phenanthrene uptake than the inherent SOM. The distribution coefficients of phenanthrene between water and the sorbed DOM on soil solids (K(ph/soc)) were about 2-3 magnitude larger than K(d) between water and inherent SOM, which may be the dominant mechanism of the enhanced sorption of phenanthrene by soils with the addition of heavy metals.     

Impact of soil management and two botanical insecticides on urease and invertase activity

Yard waste compost provides an organic amendment useful for improving soil structure and nutrient status. The activities of the enzymes hydrolyzing urea (urease) and sucrose (invertase) in the rhizosphere of potato plants were determined under field conditions. Soil urease and invertase activities were monitored in compost amended soil, grass buffer strips, and in adjacent bare soils during 35 d following soil treatment. Soil urease activity was increased by application of yard waste compost compared to untreated soil which provide evidence of increased soil microbial population following application of compost. Some transitional effects on urease activities were observed following Pyrethrins and Neemix-4E application, these effects were neither drastic nor prolonged enough to be considered deleterious to the soil microorganisms and their activities important to soil fertility. No relationship was found between invertase activity and the three management practices or soil organic matter content. It is suggested that application of botanical insecticides like pyrethrins and Neemix-4E may be useful in delaying urea fertilizer mineralization to maintain N in a form less leachable, so that the duration of N availability to plants is prolonged. The present study may also provide information on urease activity as a sensitive bioindicator of soil quality that reflects the effects of land management on soil quality and may assist land managers in monitoring long-term productivity and sustainability of agricultural lands.     

Effects of EDTA on solubility of cadmium, zinc, and lead and their uptake by rainbow pink and vetiver grass

Rainbow pink (Dianthus chinensis), a potential phytoextraction plant, can accumulate high concentrations of Cd from contaminated soils. Vetiver grass (Vetiver zizanioides) has strong and long root tissues and is a potential phytostabilization plant since it can tolerate and grow well in soils contaminated with multiple heavy metals. Soil was moderately artificially contaminated by cadmium (20 mg/kg), zinc (500 mg/kg), and lead (1000 mg/kg) in pot experiments. Three concentrations of Na2-EDTA solution (0, 5, and 10 mmol/kg soil) were added to the contaminated soils to study the influence of EDTA solution on phytoextraction by rainbow pink or phytostabilization by vetiver grass. The results showed that the concentrations of Cd, Zn, and Pb in a soil solution of rainbow pink significantly increased following the addition of EDTA (p < 0.05). The concentrations of Cd and Pb in the shoots of rainbow pink also significantly increased after EDTA solution was applied (p < 0.05), but the increase for Zn was insignificant. EDTA treatment significantly increased the total uptake of Pb in the shoot, over that obtained with the control treatment (p < 0.001), but it did not significantly increase the total uptake of Cd and Zn. The concentrations of Zn and Pb in the shoots of rainbow pink are significantly correlated with those in the soil solution, but no relationship exists with concentrations in vetiver grass. The toxicity of highly contaminating metals did not affect the growth of vetiver grass, which was found to grow very well in this study. Results of this study indicate that rainbow pink can be considered to be a potential phytoextraction plant for removing Cd or Zn from metal-contaminated soils, and that vetiver grass can be regarded as a potential phytostabilization plant that can be grown in a site contaminated with multiple heavy metals.     

Urease inhibition: a tool for toxicity identification in sediment elutriates

A method for the detection and confirmation of heavy metal toxicants in sediment elutriates based on a urease inhibition assay, ICP-AES analysis and EDTA chelation in the frame of toxicity identification evaluation (TIE) is presented. Zinc was identified as the major toxicant in pHstat elutriates of sediments of the river Saale (Germany). Implications of natural and anthropogenic chelating agents, which are frequently present in environmental samples, on toxicity confirmation of heavy metals based on the toxic unit approach are discussed.     

Determination and evaluation of cadmium, lead and nickel in greenhouse soils of Almería (Spain)

This study determines total levels of three (Cd, Pb and Ni) potentially toxic trace elements in western Almería (Spain) greenhouse surface soil horizons using microwave digestion; it establishes the geochemical baseline concentration, and it investigates possible relationships between soil properties and elemental concentrations. The results show that the soil concentration of these heavy metals is lower than mentioned in the European and Spanish normative, but they are higher than those reported by other authors working on agricultural soils. The obtained geochemical baseline concentrations (mg kg(-1)) were: Cd 0.4-0.8, Pb 2.5-89.9 and Ni 16.1-30.7. Using the upper baseline criterion, 88% of greenhouse soils have relatively higher content of heavy metals because of their Cd, Pb and Ni concentration. Moreover, soil properties are related to heavy metals contents suggesting that among Cd, Pb and Ni have a similar origin and those total metal concentrations are controlled primarily by soil compositions.     

功能微生物对污染农田土壤中铅锌的溶出实验

用微生物修复重金属污染的土壤是当今的研究热点之一。试图寻找一类可以增加土壤中重金属铅、锌的可溶态的功能微生物,并研究该微生物在不同土壤含水率和不同投菌比条件下对铅、锌溶出作用的影响。最终筛选出符合条件的菌JK3,并发现当土壤含水率为35%,投菌比为0.15 mL/g土(菌液OD=1)时,土壤中铅和锌的可溶态增加量最多。其中,可溶态铅的含量由原来的0.49 mg/kg增加到了5.04 mg/kg,可溶态锌的含量由原来的2.23 mg/kg增加到了22.44mg/kg。该方法为后续用植物提取或化学淋洗等方法将土壤中的重金属彻底去除提供了可能。     

Impact of Soil Management and Two Botanical Insecticides on Urease and Invertase Activity

Yard waste compost provides an organic amendment useful for improving soil structure and nutrient status. The activities of the enzymes hydrolyzing urea (urease) and sucrose (invertase) in the rhizosphere of potato plants were determined under field conditions. Soil urease and invertase activities were monitored in compost amended soil, grass buffer strips, and in adjacent bare soils during 35 d following soil treatment. Soil urease activity was increased by application of yard waste compost compared to untreated soil which provide evidence of increased soil microbial population following application of compost. Some transitional effects on urease activities were observed following Pyrethrins and Neemix‐4E application, these effects were neither drastic nor prolonged enough to be considered deleterious to the soil microorganisms and their activities important to soil fertility. No relationship was found between invertase activity and the three management practices or soil organic matter content. It is suggested that application of botanical insecticides like pyrethrins and Neemix‐4E may be useful in delaying urea fertilizer mineralization to maintain N in a form less leachable, so that the duration of N availability to plants is prolonged. The present study may also provide information on urease activity as a sensitive bioindicator of soil quality that reflects the effects of land management on soil quality and may assist land managers in monitoring long‐term productivity and sustainability of agricultural lands.     

Interactions between arbuscular mycorrhizae and heavy metals under sand culture experiment

A sand culture experiment was established to determine interactions between arbuscular mycorrhizae and heavy metals. Mycorrhizal infection rates, spore densities, maize root and shoot weights, and heavy metal contents in maize were as indexes of responses of arbuscular mycorrhizal fungi (Acaulospora laevis, Glomus caledonium and Glomus manihotis) to heavy metals (Cu and Cd). The mycorrhizal infection rates of G. caledonium were the highest among these three mycorrhizal fungi, but the sporulating ability of G. caledonium was the poorest in the heavy metal treatments. The shoot and root weights of non-mycorrhizal plants were usually greater than those of mycorrhizal plants when the Cu concentrations in solutions are less than 3 mg l(-1) or Cd concentrations less than 1 mg l(-1). When Cd concentrations were 0.5 and 1 mg(-1), the root and shoot weights of plants inoculated with A. laevis were significantly (p < 0.05) lower than those of other treatments. Copper concentrations in shoots of mycorrhizal plants were higher than those of non-mycorrhizal ones at all Cu concentrations in solution, especially at low Cu concentrations. As to A. laevis, Cu concentrations in roots and shoots of the host were higher than those of non-mycorrhizal plants in these treatments. Thus A. laevis was sensitive to Cu and Cd, especially Cd, and G. caledonium was more tolerant to these two heavy metals. It is suggested that G. caledonium might be a promising mycorrhizal fungus for bioremediation of heavy metal contaminated soil.     

Influence of organic solvents on the sensitivity of a bioluminescence toxicity test with Vibrio harveyi

The possibility of using organic solvents (OSs) to increase the susceptibility of bioluminescent microorganisms in a bioassay for assessing the toxicity of chemicals dissolved in water was investigated. To conduct the tests acetonitrile, dimethyl sulfoxide, ethanol, methanol, and isopropanol were used as OSs and Cd, Hg, and Zn as reference toxicants. The addition of OSs modified the toxicity of the three metals to Vibrio harveyi, according on the bioluminescence assay used. The sensitivity of the luminescence bioassay for Hg increased in the presence of the five OSs, thus indicating a greater toxic effect. However, the sensitivity of the assay for the other two metals, Cd and Zn, increased or decreased (lesser toxic effect) depending on the concentration at which the OSs were used. No correlation was observed between the concentration of the five OSs and the toxicity of the three reference toxicants. From this it can be deduced that none of these OSs could be recommended for increasing generically the sensitivity of toxicity biotests using V. harveyi.     

外源Cd对不同利用方式红壤脲酶活性的影响

采用室内培养实验,通过将外源Cd添加到同一母质、全镉含量相近的不同利用方式的红壤中(林地、水稻土和菜园土),研究了外源Cd污染对不同利用方式红壤脲酶活性的影响。结果表明:整个培养过程中,Cd污染对3种红壤脲酶活性都有抑制作用,且随重金属浓度的增强而增强。同剂量Cd污染对3种红壤脲酶活性的抑制效应不同,大小为林地>水稻土>菜园土。实验设定的Cd处理水平下,对林地、水稻土和菜园土脲酶活性产生显著抑制作用(p<0.05)的Cd浓度分别为5、30和50 mg/kg土。     

微生物矿化修复重金属污染土壤

以选矿厂附近土壤为研究对象,分析了土壤中交换态重金属含量,As、Pb、Cd、Zn和Cu的交换态浓度为14.01、4.95、0.64、33.46和12.95 mg/kg。基于生物矿化原理,利用碳酸盐矿化菌生长代谢过程产生的脲酶来分解底物尿素,产生碳酸根离子,固结重金属离子,使得土壤中活泼的重金属离子转变为碳酸盐矿物态,降低其危险。研究了温度、pH和重金属离子对酶活性的影响,发现环境30℃温度有利于促进酶活性;在弱酸性条件下,底物分解量减少15%;重金属离子在低浓度时对脲酶活性影响不大,浓度提高后对酶活性抑制作用没有加剧。将制备好的微生物矿化修复制剂喷洒于1 000 m2的污染土壤中,实验结果发现,土壤中交换态重金属离子含量在0～20 cm范围内明显减少,As、Pb、Cd、Zn和Cu的交换态浓度分别减少至2.37、1.25、0.31、16.67和3.42 mg/kg。     

Effects of Metsulfuron-Methyl on the Microbial Population and Enzyme Activities in Wheat Rhizosphere Soil

The effects of metsulfuron-methyl, a sulfonylurea herbicide, on the wheat soil microorganisms were evaluated by the methods of microbial inoculation culture, and the activities of three enzymes were measured using the colorimetric method. The tolerant microorganisms that can resist 500 μ g·g^?1 metsulfuron-methyl in the counting culture medium were studied specially. Metsulfuron-methyl distinctly inhibited the common aerobic heterotriphic bacteria, but the effects on common fungi and common actinomycete were not evident. In the meantime, the number of tolerant fungi increased greatly in the rhizosphere after the application of metsulfuron-methyl in contrast to the significant decrease of the amount of tolerant actinomycete. It indicates that fungi might turn into the dominant microbial type and actinomycete is the sensitive factor in the soil polluted by sulfonylurea residues. The population of aromatic compounds–decomposing bacteria, aerobic azotobacter, and nitrite bacteria all increased in the earlier period, but the aerobic azotobacter decreased rapidly in number 30 days later, and the amount of nitrite bacteria also showed a temporary decrease with time 15 days later. However, the denitrifying bacteria just began to increase significantly after the crops had grown for 50 days. The amount of sulfur-oxidizing bacteria gradually decreased with the growth of crops, and so were the sulfate-reducing bacteria after metsulfuron-methyl application. To all types of microorganisms, there were more microbes in rhizosphere samples than those in nonrhizosphere except aerobic azotobacter. It means the growth of wheat root system can stimulate the growth of most microorganisms. The activities of hydrogen peroxidase and polyphenol oxidase in soil samples after metsulfuron-methyl application were notably lower than those in the control, and the difference of the activities between the samples of rhizosphere and nonrhizosphere was evident. On the contrary, the activity of dehydrogenase was not inhibited by the application of metsulfuron-methyl, and the rhizosphere effect was not obvious either.     

Effects of metsulfuron-methyl on the microbial population and enzyme activities in wheat rhizosphere soil

The effects of metsulfuron-methyl, a sulfonylurea herbicide, on the wheat soil microorganisms were evaluated by the methods of microbial inoculation culture, and the activities of three enzymes were measured using the colorimetric method. The tolerant microorganisms that can resist 500 microg x g(-1) metsulfuron-methyl in the counting culture medium were studied specially. Metsulfuron-methyl distinctly inhibited the common aerobic heterotriphic bacteria, but the effects on common fungi and common actinomycete were not evident. In the meantime, the number of tolerant fungi increased greatly in the rhizosphere after the application of metsulfuron-methyl in contrast to the significant decrease of the amount of tolerant actinomycete. It indicates that fungi might turn into the dominant microbial type and actinomycete is the sensitive factor in the soil polluted by sulfonylurea residues. The population of aromatic compounds-decomposing bacteria, aerobic azotobacter, and nitrite bacteria all increased in the earlier period, but the aerobic azotobacter decreased rapidly in number 30 days later, and the amount of nitrite bacteria also showed a temporary decrease with time 15 days later. However, the denitrifying bacteria just began to increase significantly after the crops had grown for 50 days. The amount of sulfur-oxidizing bacteria gradually decreased with the growth of crops, and so were the sulfate-reducing bacteria after metsulfuron-methyl application. To all types of microorganisms, there were more microbes in rhizosphere samples than those in nonrhizosphere except aerobic azotobacter. It means the growth of wheat root system can stimulate the growth of most microorganisms. The activities of hydrogen peroxidase and polyphenol oxidase in soil samples after metsulfuron-methyl application were notably lower than those in the control, and the difference of the activities between the samples of rhizosphere and nonrhizosphere was evident. On the contrary, the activity of dehydrogenase was not inhibited by the application of metsulfuron-methyl, and the rhizosphere effect was not obvious either.     

雷州半岛土壤重金属分布特征及其污染评价

在雷州半岛采集了106个土壤表层样品,分析了其中8种重金属元素(Cu、Pb、Zn、Cr、Ni、Cd、Hg和As)的全量.结果表明,雷州半岛土壤重金属污染由高到低排序为Ni＞Cr＞Hg＞Cu＞Zn＞Cd＞As＞Pb,Zn、Cd、As和Pb质量浓度均没有超标,Hg和Cu质量浓度超标率亦不高,但Ni和Cr平均质量浓度达49.81、87.13 mg/kg,高于国内外其他对照区域,超标率分别为25.47%和24.53%;重金属元素在雷州半岛各土壤利用类型中分布规律不明显,按4种主要土壤利用类型受重金属污染程度大小排序为甘蔗地＞果园土＞水田＞菜地;雷州半岛土壤综合污染指数总平均为0.970,土壤总体上尚清洁,重金属污染处于警戒水平;雷州半岛各区域中,徐闻、雷州两地土壤重金属质量浓度明显高于其他地区,其主要原因是徐闻、雷州两地成土母质主要为玄武岩,造成土壤Cr、Ni及其他重金属背景值较高.     

Effect of applying chemical fertilizers on forms of lead and cadmium in red soil

A three-month incubation study was undertaken to examine the influence of N, P and K on the various forms (soluble plus exchangeable (SE), weakly specifically adsorbed (WSA), Fe-Mn oxides bound (OX), organic matter complexed (OM) and residual fractions (RES)) of lead (Pb) and cadmium (Cd) in a red soil. Application of urea at the rate of 200 mg N/kg significantly lowered the SE fraction, but raised the WSA or OX fraction of both metals. Supply of 80 mg P/kg caused a decrease in the SE fraction of the two metals. The WSA fraction of Pb was reduced, whereas that of Cd increased by adding P. However, addition of 100 mg K/kg led to an increase in the SE fraction, but a decrease in the WSA fraction of Pb and Cd. Applying chemical fertilizers had no significant consistent influences on the other fractions of metals. These findings suggest that in heavy metal contaminated red soil, applying fertilizers does not only provide plant nutrients, but may also change the speciations and thus biovailability of heavy metals.     

EDTA强化电动力学修复重金属复合污染土壤

在自制的电动力学装置中,研究多种重金属复合污染土壤的电动力学修复,通过在阴极添加络合剂EDTA来提高修复效率。实验结果表明,EDTA的引入提高了修复过程中的电流值,且EDTA与重金属的络合提高了污染物向电极液的迁移效率,从而强化了电动力学修复效果。在设定的浓度(0、0.01、0.02、0.05和0.1 mol/L)中,0.1 mol/L的EDTA具有最佳的修复效率。在此实验条件下,污染土壤中的总铜、总铅和总镉的去除率分别为90.2%、68.1%和95.1%。电动力学修复后,对土壤重金属进行化学形态分析,发现电动力学修复显著改变了土壤重金属存在形态,修复后土壤中的铜、铅、镉主要以较稳定的有机态和残余态形式存在,显著降低了对周边生物和环境的毒害。     

重金属污染土壤接种丛枝菌根真菌对蚕豆毒性的影响

采用盆栽实验的方法,研究了重金属(包括Cu、Zn、Pb和Cd)复合污染和接种丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)Glomus mosseae对蚕豆(Vicia faba)生长及DNA损伤的影响.结果表明,虽然接种菌根真菌对蚕豆生物量的影响并不显著,但是却显著影响植物对重金属的吸收,接种菌根真菌对蚕豆吸收4种重金属元素的作用有差异.采用单细胞凝胶电泳(single cell gel electrophoresis,SCGE)法研究接种菌根真菌对蚕豆叶片的DNA损伤的影响,与重金属吸收的结果相吻合.结果表明,接种处理可显著增加蚕豆叶片的DNA损伤程度,这与接种处理可提高植物的重金属吸收相一致.