Historical alteration in the nitrogen concentration and N natural abundance of mosses in Germany: Indication for regionally varying changes in atmospheric nitrogen deposition within the last 140 years

Historical alterations of nitrogen deposition in the western part of Germany were investigated by comparing nitrogen concentrations and ¹⁵N natural abundance of historical and recent samples of the two pleurocarpous mosses Pleurozium schreberi and Scleropodium purum. Pooling of the data revealed only slight tissue N increases over the past 140 years which were significant nevertheless. At closer examination on the single site level historical increases of N concentrations were found particularly for some sites in regions where agricultural activities were considerably intensified during the second half of the 20th century. The comparison of δ¹⁵N values showed a strong depletion of ¹⁵N natural abundance in areas currently heavily influenced by livestock management. This indicates an increased impact of NH_y compounds. However, the almost unchanged δ¹⁵N values in some low mountain range areas with only moderate intensification of agriculture point to a more or less constant ratio of NH_y/NO_x input over time. Significant correlations of both tissue N concentrations and ¹⁵N natural abundance between the two species justify the assumption that they use the same nitrogen source, probably atmospheric deposition.     

Assessment of the quality of the air in the city of Palermo through chemical and cell analyses on Pinus needles

The influence of air pollution on the chemical composition of Pinus sp. needles was examined in polluted and control sites in and around the city of Palermo (Sicily). The chemical composition of needles indicated the extent of contamination of the trees, which were cytologically examined. Cell analysis was carried out on pine samples, including needles and pollens, from 15 different locations. Biostructural and spectrophotometric tests were performed. In particular, concentrations of toxic (Cd, Pb) and non-toxic metals (Fe,Cu, Zn) were determined, as well as injury caused by their accumulation in the needles. The more highly urbanised areas showed higher concentrations of metals (Pb, Cu. Zn, Fe); only the concentrations of Cd and Mn turned out to be constant in all the sites. Cell analysis revealed displasic cells and secondary metabolite accumulations in trees from polluted sites. These changes observed were most likely caused by the toxic effect of pollutants.     

Euglena gracilis as a model for the study of Cu and Zn toxicity and accumulation in eukaryotic cells

We have observed the effect of copper and zinc on the biology of Euglena gracilis. The cells displayed different sensitivities to these metals, as the apparent LC₅₀ for Cu²⁺ was 0.22 mM, and for Zn²⁺ it was 0.88 mM. While Zn²⁺ was able to increase cell proliferation even at 0.1 mM, the minimal CuCl₂ concentration tested (0.02 mM) was sufficient to impair cell division. Higher concentrations of these metals not only inhibited cell division in a concentration-dependent manner, but also interfered with the metabolism of E. gracilis. A higher accumulation of proteins and lipids per cell was observed at the DI₅₀ concentration for metal-treated cells. These results suggest that the test concentration of both metals leads to a failure in completing cell division. Ultrastructural analysis indicated a chloroplast disorganization in copper-treated cells, as well as the presence of electron dense granules with different shapes and sizes inside vacuoles. Microanalysis of these granules indicated an accumulation of copper, thus suggesting a detoxification role played by the vacuoles. These results indicate that E. gracilis is an efficient biological model for the study of metal poisoning in eukaryotic cells. They also indicate that copper and zinc (copper being more poisonous) had an overall toxic effect on E. gracilis and that part of the effect can be ascribed to defects in the structure of chloroplast membranes.     

Surfactant-based oil dispersant toxicity to developing nauplii of Artemia: effects on ATPase enzymatic system

The paper deals with the toxicity of a surfactant-based oil dispersant to the ATPase activities of two naupliar stages of Artemia (instar I & II). Both instars were exposed to sub-lethal and lethal concentrations derived from acute toxicity data. The chosen concentrations were near to LOECs and NOECs. An eightfold difference indicated between the instars was instar-exposure time dependent. The most prominent effects were the inhibition and the stimulation of Na⁺/K⁺-ATPase and Mg²⁺-ATPase activities, respectively. The cause of these effects was related to the dispersant components, the surfactants. The pattern stimulation/inhibition of Mg²⁺-ATPase and Na⁺/K⁺-ATPase activities could be used to indicate toxic stress by surfactant-based oil dispersants since previous studies with other contaminants have shown different ATPase activity patterns.     

污泥堆肥施用于草坪草栽培:Zn和Cu的分布特征简

为了探索污泥堆肥中重金属在土壤-植物系统中的积累与转移特性,通过温室盆栽实验,分析了污泥堆肥对草坪草高羊茅、黑麦草和白三叶生物量积累的情况,研究了污泥堆肥中Zn和Cu在植物和土壤中的分布特征。结果表明,污泥堆肥施用可以有效促进3种草坪草的积累生物量,在0~6 kg/m²的污泥堆肥施用量范围内,草坪草的生物量积累随着施用量的增加而提高。土壤中Zn和Cu的含量随污泥堆肥施加量的增加而增大,85%以上的Zn和Cu残留在土壤中。污泥堆肥中的Zn和Cu均可以被植物吸收,随着污泥堆肥施用量的增加,草坪草对Zn和Cu的吸收量增大,但当污泥堆肥施用量超过一定阈值时,草坪草吸收Zn和Cu不再增加,甚至减少;对于不同的草坪草,这一阈值有所不同。植物对Zn和Cu的吸收量只占土壤中Zn和Cu减少量的5%左右。根据生物富集系数（BCF）的计算结果推测,污泥堆肥的施用对土壤环境的影响大于对植物体内累积Zn和Cu的影响。     

Cd、Ni单一及复合污染对土壤酶活性的影响

采用外源添加重金属和露天盆栽实验研究了Cd、Ni对含羞草、三叶草根际土壤脲酶、过氧化氢酶和蔗糖酶活性的影响。结果表明,Cd、Ni单一污染条件下,低浓度的Cd、Ni对含羞草组、三叶草组的脲酶、过氧化氢酶有激活作用,高浓度的Cd、Ni对土壤脲酶、过氧化氢酶有一定的抑制作用;对蔗糖酶有较强的抑制作用。在Cd、Ni复合污染条件下,对含羞草组、三叶草组的脲酶、过氧化氢酶和蔗糖酶产生抑制作用;Cd、Ni单一及复合污染对土壤酶活性的抑制大小顺序为:脲酶>蔗糖酶>过氧化氢酶,其中土壤脲酶可以作为Cd、Ni污染的预警指标。含羞草能显著提高土壤脲酶、过氧化氢酶和蔗糖酶的活性,修复能力大于三叶草,在Cd、Ni污染修复方面有很好的应用前景。     

Reed beds receiving industrial sludge containing nitroaromatic compounds

Goal, Scope and Background Sweden has prohibited the deposition of organic waste since January, 2005. Since 1 million tons of sludge is produced every year in Sweden and the capacity for incineration does not fill the demands, other methods of sludge management have to be introduced to a larger degree. One common method in the USA and parts of Europe is the use of wetlands to treat wastewater and sewage sludge. The capacity of reed beds to affect the toxicity of a complex mixture of nitroaromatics in sludge, however, is not fully elucidated. In this study, an industrial sludge containing explosives and pharmaceutical residues was therefore treated in artificial reed beds and the change in toxicity was studied. Nitroaromatic compounds, which are the main ingredients of many pharmaceuticals and explosives, are well known to cause cytotoxicity and genotoxicity. Recently performed studies have also showed that embryos of zebrafish (Danio rerio) are sensitive to nitroaromatic compounds. Therefore, we tested the sludge passing through constructed wetlands in order to detect any changes in levels of embryotoxicity, genotoxicity and dioxin-like activity (AhR-agonists). We also compared unplanted and planted systems in order to examine the impact of the root system on the fate of the toxicants. Methods An industrial sludge containing a complex mixture of nitroaromatics was added daily to small-scale constructed wetlands (vertical flow), both unplanted and planted with Phragmites australis. Sludge with an average dry weight of 1.25%, was added with an average hydraulic loading rate of 1.2 L/day. Outgoing water was collected daily and stored at −20°C. The artificial wetland sediment was Soxhlet extracted, followed by clean-up with multi-layer silica, or extracted by ultrasonic treatment, yielding one organic extract and one water extract of the same sample. Genotoxicity of the extracts was measured according to the ISO protocol for the umu-C genotoxicity assay (ISO/TC 147/SC 5/WG9 N8), using Salmonella typhimurium TA1535/pSK1002 as test organism. Embryotoxicity and teratogenicity were studied using the fish egg assay with zebrafish (Danio rerio) and the dioxin-like activity was measured using the DR-CALUX assay. Chemical analyses of nitroaromatic compounds were performed using Solid Phase Micro Extraction (SPME) and GC-MS. Results Organic extracts of the bed material showed toxic potential in all three toxicity tests after two years of sludge loading. There was a difference between the planted and the unplanted beds, where the toxicity of organic extracts overall was higher in the bed material from the planted beds. The higher toxicity of the planted beds could have been caused by the higher levels of total carbon in the planted beds, which binds organic toxicants, and by enrichment caused by lower volumes of outgoing water from the planted beds. Discussion Developmental disorders were observed in zebrafish exposed directly in contact to bed material from unplanted beds, but not in fish exposed to bed material from planted beds. Hatching rates were slightly lower in zebrafish exposed to outgoing water from unplanted beds than in embryos exposed to outgoing water from planted beds. Genotoxicity in the outgoing water was below detection limit for both planted and unplanted beds. Most of the added toxicants via the sludge were unaccounted for in the outgoing water, suggesting that the beds had toxicant removal potential, although the mechanisms behind this remain unknown. Conclusions During the experimental period, the beds received a sludge volume (dry weight) of around three times their own volume. In spite of this, the toxicity in the bed material was lower than in the sludge. Thus, the beds were probably able to actually decrease the toxicity of the added, sludge-associated toxicants. When testing the acetone extracts of the bed material, the planted bed showed a higher toxicity than the unplanted beds in all three toxicity tests. The toxicity of water extracts from the unplanted beds, detected by the fish egg assay, were higher than the water extracts from the planted beds. No genotoxicity was detected in outgoing water from either planted or unplanted beds. All this together indicates that the planted reed beds retained semi-lipophilic acetone-soluble toxic compounds from the sludge better than the unplanted beds, which tended to leak out more of the water soluble toxic compounds in the outgoing water. The compounds identified by SPME/GC in the outgoing water were not in sufficient concentrations to have caused induction in the genotoxicity test. Recommendations and Perspectives This study has pointed out the benefits of using constructed wetlands receiving an industrial sludge containing a complex mixture of nitroaromatics to reduce toxicity in the outgoing water. The water from planted, constructed wetlands could therefore be directed to a recipient without further cleaning. The bed material should be investigated over a longer period of time in order to evaluate potential accumulation and leakage prior to proper usage or storage. The plants should be investigated in order to examine uptake and possible release when the plant biomass is degraded. This article has been developed on the basis of a presentation given at the Annual meeting of SETAC Europe German Language Branch 2004 in Aachen. ESS-Submission Editor: Dr. Ludek Blaha (blaha@recetox.muni.cz)     

Phytoremediation potential of the novel atrazine tolerant Lolium multiflorum and studies on the mechanisms involved

Atrazine impact on human health and the environment have been extensively studied. Phytoremediation emerged as a low cost, environmental friendly biotechnological solution for atrazine pollution in soil and water. In vitro atrazine tolerance assays were performed and Lolium multiflorum was found as a novel tolerant species, able to germinate and grow in the presence of 1 mg kg⁻¹ of the herbicide. L. multiflorum presented 20% higher atrazine removal capacity than the natural attenuation, with high initial degradation rate in microcosms. The mechanisms involved in atrazine tolerance such as mutation in psbA gene, enzymatic detoxification via P₄₅₀ or chemical hydrolysis through benzoxazinones were evaluated. It was demonstrated that atrazine tolerance is conferred by enhanced enzymatic detoxification via P₄₅₀. Due to its atrazine degradation capacity in soil and its agronomical properties, L. multiflorum is a candidate for designing phytoremediation strategies for atrazine contaminated agricultural soils, especially those involving run-off avoiding.     

地表水中隐孢子虫和贾第鞭毛虫微球的混凝去除

以向地表水中投加的隐孢子虫微球和贾第鞭毛虫微球为对象,研究通过对给水混凝工艺对其的去除效果进行比较,着重考察了不同混凝剂种类、投加量及水样pH值对去除效果的影响.结果表明,混凝工艺可较为有效地去除地表水中的两虫微球,去除率最高可达4 lg.相比常规的金属盐类混凝剂,高分子的PAC、PAFC混凝剂对两虫微球的混凝去除率更...     

The arbuscular mycorrhizal fungus Glomus mosseae can enhance arsenic tolerance in Medicago truncatula by increasing plant phosphorus status and restricting arsenate uptake

A pot experiment examined the biomass and As uptake of Medicago truncatula colonized by the arbuscular mycorrhizal (AM) fungus Glomus mosseae in low-P soil experimentally contaminated with different levels of arsenate. The biomass of G. mosseae external mycelium was unaffected by the highest addition level of As studied (200 mg kg⁻¹) but shoot and root biomass declined in both mycorrhizal and non-mycorrhizal plants, indicating that the AM fungus was more tolerant than M. truncatula to arsenate. Mycorrhizal inoculation increased shoot and root dry weights by enhancing host plant P nutrition and lowering shoot and root As concentrations compared with uninoculated plants. The AM fungus may have been highly tolerant to As and conferred enhanced tolerance to arsenate on the host plant by enhancing P nutrition and restricting root As uptake.     

大型蚤标志酶用于城市二级出水毒性的评价

大型蚤是一种国际公认的标准实验生物,广泛应用于污水、地表水等水质毒性检测。毒性水平较低的城市二级出水,对大型蚤往往无急性毒性效应,而具有慢性毒性效应,但慢性毒性检测周期过长,因此探索一种更灵敏的指标,实现快速检测,对于控制二级出水的水质风险具有重要意义。本研究考察了大型蚤在短期暴露于城市二级出水条件下,其体内乙酰胆碱酯酶、超氧化物歧化酶、过氧化氢酶、ATP酶、羧酸酯酶和碱性磷酸酯酶的酶活变化特征,从中筛选出对二级出水毒性响应灵敏的标志酶指标。实验结果表明,碱性磷酸酯酶、过氧化氢酶对二级出水毒性响应相对较灵敏,具有成为标志酶的潜力,研究结果为城市二级出水生物毒性评价方法优化提供新的思路。     

Fungal hydroxylation of polychlorinated naphthalenes with chlorine migration by wood rotting fungi

Biodegradation of the polychlorinated naphthalenes (PCNs) 1,4-dichloronaphthalene (1,4-DCN), 2,7-dichloronaphthalene (2,7-DCN), and 1,2,3,4-tetrachloronaphthalene (1,2,3,4-TCN), by the white-rot fungus Phlebia lindtneri was investigated. 1,4-DCN was metabolized to form six metabolites by the fungus. It was estimated from GC–MS fragment patterns that the metabolites were four putative hydroxylated and two dihydrodihydroxylated compounds. One of the hydroxylated products was identified as 2,4-dichloro-1-naphthol by GC–MS analysis using an authentic standard. This intermediate indicated chlorine migration in a biological system of P. lindtneri. 2,7-DCN was metabolized to five hydroxylated metabolites and a dihydrodihydroxylated metabolite. Significant inhibition of the degradation of DCNs and formation of their metabolic products was observed in incubation with the cytochrome P-450 monooxygenase inhibitor piperonyl butoxide. The formation of the dihydrodiol-like metabolites, chlorine migration and the experiment with P-450 inhibitor suggested that P. lindtneri provides hydroxyl metabolites via benzene oxide intermediates of DCNs by a cytochrome P450 monooxygenase. In addition, P. lindtneri degraded 1,2,3,4-TCN; two hydroxylated compounds and a dihydrodihydroxylated compound were formed.     

不同质量浓度黄菖蒲和狭叶香蒲对铜绿微囊藻的化感作用简

分别采用黄菖蒲（Iris pseudacorus L.）、狭叶香蒲（Typha angustifolia L.）和铜绿微囊藻（Microcystis aeruginosa）共生培养的实验方法研究了不同质量浓度黄菖蒲、狭叶香蒲对铜绿微囊藻的化感作用。结果表明,黄菖蒲在质量浓度大于10 g/L时对初始密度为1.0×10⁷ ind/mL的铜绿微囊藻具有较好的抑制作用,表现为黄菖蒲质量浓度为10、20和40 g/L时,第15天对铜绿微囊藻的抑制率分别为30.1%、51.8%和84.0%;狭叶香蒲在质量浓度大于20 g/L时对铜绿微囊藻有明显的抑制作用,表现为狭叶香蒲质量浓度为20 g/L和40 g/L时,第15天对铜绿微囊藻的抑制率分别为34.2%和77.7%,实验过程中,铜绿微囊藻叶绿素a含量逐渐减少,而藻密度、SOD活性及MDA含量先增加后逐渐降低,表明经过一段时间持续地化感胁迫,黄菖蒲和狭叶香蒲可以诱导铜绿微囊藻产生氧化胁迫,导致细胞结构严重损伤和叶绿素大量分解,从而强烈抑制铜绿微囊藻的生长。     

全氟辛烷磺酸盐(PFOS)在藻渣/小球藻上的吸附行为及机理

为了开发新型廉价生物吸附剂,以高效吸附去除水体中全氟辛烷磺酸盐(PFOS),对小球藻提取生物柴油后的藻渣吸附酸性水体中的PFOS进行了吸附行为及机理的研究。小球藻提取生物质柴油后,比表面积、孔容、孔径几乎没有变化;等电点由3.3降低至2.7;蛋白质含量由51.45%提高到57.35%。在酸性条件下(pH≤3),小球藻和藻渣对PFOS的吸附率均达到99%以上;随着pH值增加至7,二者的吸附去除率迅速降低,但仍保持在22%~26%。小球藻和藻渣对PFOS的最大吸附容量分别为353.69 mg/g和444.83 mg/g。Freundlich模型能较好地拟合二者对PFOS的吸附数据,表明为多层吸附,即小球藻以静电吸引的形式吸附PFOS阴离子,并疏水分配至所含蛋白质中;而藻渣中含量较高的蛋白质对PFOS的疏水性分配作用是导致藻渣吸附量增高的主要原因。     

Toxicity of cobalt-complexed cyanide to <Emphasis Type="Italic">Oncorhynchus mykiss,Daphnia magna</Emphasis>, and <Emphasis Type="Italic">Ceriodaphnia dubia</Emphasis>

Background Cobalt cyanide complexes often result when ore is treated with cyanide solutions to extract gold and other metals. These have recently been discovered in low but significant concentrations in effluents from gold leach operations. This study was conducted to determine the potential toxicity of cobalt-cyanide complexes to freshwater organisms and the extent to which ultraviolet radiation (UV) potentiates this toxicity. Tests were also conducted to determine if humic acids or if adaptation to UV influenced sensitivity to the cyanide complexes. Methods Rainbow trout (Oncorhynchus mykiss), Daphnia magna, and Ceriodaphnia dubia were exposed to potassium hexacyanocobaltate in the presence and absence of UV radiation, in the presence and absence of humic acids. Cyano-cobalt exposures were also conducted with C. dubia from cultures adapted to elevated UV. Results With an LC50 concentration of 0.38 mg/L, cyanocobalt was over a 1000 times more toxic to rainbow trout in the presence of UV at a low, environmentally relevant irradiance level (4 μW/cm² as UVB) than exposure to this compound in the absence of UV with an LC50 of 112.9 mg/L. Toxicity was immediately apparent, with mortality occurring within an hour of the onset of exposure at the highest concentration. Fish were unaffected by exposure to UV alone. Weak-acid dissociable cyanide concentrations were observed in irradiated aqueous solutions of cyanocobaltate within hours of UV exposure and persisted in the presence of UV for at least 96 hours, whereas negligible concentrations were observed in the absence of UV. The presence of humic acids significantly diminished cyanocobalt toxicity to D. magna and reduced mortality from UV exposure. Humic acids did not significantly influence survival among C. dubia. C. dubia from UV-adapted populations were less sensitive to metallocyanide compounds than organisms from unadapted populations. Conclusions The results indicate that metallocyanide complexes may pose a hazard to aquatic life through photochemically induced processes. Factors that decrease UV exposure such as dissolved organic carbon or increased pigmentation would diminish toxicity.     

氧化亚铁硫杆菌浸铜研究及胞外多聚物的作用

为了获得氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,A.f)以及胞外多聚物(extracellular polymeric sub-stances,EPS)对铜浸出的作用结果,进行了透析袋实验,设置2个处理,2者的浸出液均为成熟菌液,但一个将铜片装入透析袋中以隔离A.f及EPS与铜的直接接触。结果表明:(1)菌液的处理和加透析袋的处理中铜都得到了浸出,240 h铜的浸出浓度分别为6 796 mg/L和1 366 mg/L,可知A.f及EPS与铜片直接接触能促进铜的浸出;(2)透析袋的处理中Fe3+保持较高浓度,说明Fe3+与EPS络合后被阻挡在透析袋外面从而不能与铜反应,可知EPS在A.f浸铜中起了媒介载体的作用;(3)2者的SEM图表明无透析袋的处理A.f与铜在实验初期发生了吸附,实验后期则无,实验初期铜快速浸出是由于传质距离短,而后期速度变缓是由于传质距离增大。     

Combination of in vitro bioassays for the determination of cytotoxic and genotoxic potential of wastewater, surface water and drinking water samples

In this study we evaluated genotoxicity and cytotoxicity of native samples of wastewaters (15 samples), surface waters (28 samples) and potable waters (8 samples) with the SOS/umuC assay with Salmonella typhimurium TA1535/pSK1002 and MTT assay with human hepatoma HepG2 cells. The genotoxicity of selected samples was confirmed with the comet assay with HepG2 cells. In the SOS/umuC assay 13 out of the 51 samples were genotoxic: two effluent samples from chemical industry; one sample of wastewater treatment plant effluent; two hospital wastewater samples; three river water samples and four lake water samples. Six samples were cytotoxic for HepG2 cells: both effluent samples of chemical industry, two wastewater treatment plant effluent samples, and two river water samples, however, only the chemical industry effluent samples were genotoxic and cytotoxic, indicating that different contaminants are responsible for genotoxic and toxic effects. Comparing genotoxicity of river and lake water samples with the chemical analytical data of the presence of the residues of pharmaceutical and personal care products (non-steroidal anti-inflammatory drugs, UV filters and disinfectants) in these samples, indicated that the presence of UV filters might be linked to the genotoxicity of these samples. The results showed that the application of the bacterial SOS/umuC assay and mammalian cell assays (MTT and comet assay) with HepG2 cells was suitably sensitive combination of assays to monitor genotoxicity and cytotoxicity of native samples of wastewaters and surface waters. With this study we also confirmed that the toxicity/genotoxicity bioassays should be an integral tool in the evaluation of toxicity of complex wastewaters before the release into environment, as well as for the monitoring of surface water quality, providing data useful in risk assessment.     

菖蒲和空心菜在处理微污染河水潜流人工湿地中的应用

为了探讨修复微污染河水的潜流湿地中植物对污染物去除效果的影响及其生长变化,在野外条件下构建2座分别栽种菖蒲和空心菜的水平潜流人工湿地,并以未栽种植物的湿地作空白。分析了湿地中污染物的去除效果,考察了湿地中植物的生物量、根系活力和氮磷含量的变化。植物湿地中污染物净化效果优于空白湿地,菖蒲和空心菜湿地对氨氮(NH+4-N)、总氮(TN)、总磷(TP)和高锰酸盐指数(CODMn)的平均去除率分别为61.1%和57.5%,31.5%和39.7%,24.7%和25.5%,20.4%和20.7%。实验结果表明,湿地中菖蒲的根系鲜重是空心菜的4.2倍,但其根系活力低于空心菜。2种植物均可在湿地中正常生长,但受湿地中营养盐浓度的限制性影响,移栽后的植物组织氮磷含量与移栽前相比下降了11.8%~20.3%。植物在净化微污染河水的潜流人工湿地中对N、P的去除起重要作用。