Contribution for tier 1 of the ecological risk assessment of Cunha Baixa uranium mine (Central Portugal): II. soil ecotoxicological screening

This study presents the first ecotoxicological data concerning the soils of the area surrounding the Cunha Baixa uranium mine. Our main goal was to categorise soils from the area based on their toxicity profiles using a battery of cost- and time-effective bioassays (elutriate approach - Microtox and Daphnia acute tests; whole-soil approach - Microtox and avoidance assays with Eisenia andrei), as a part of tier 1 of an ongoing Environmental Risk Assessment. No acute toxicity was found for any of the 10 sites/soils using Microtox or Daphnia. On the contrary, the behavioural response of E. andrei was found to be an extremely sensitive endpoint, allowing the discrimination of highly to moderately toxic soils based on their toxicity profiles (as a function of soil concentration). Soils exhibiting highest toxicity corresponded to areas subjected to runoffs or sludge deposition from the aquatic effluent, while non-toxic soils were farthest to the mine. Data obtained in avoidance assays strengthen the previous evaluation of risks based on chemical data and supported decisions about proceeding for tier 2.     

Evaluation of wastewater toxicity: comparative study between Microtox and activated sludge oxygen uptake inhibition

Microtox is a frequent toxicity tool for the screening of wastewaters discharged into wastewater treatment plants. There is currently an increasing controversy between this test and others using activated sludge. A Microtox and electrolytic respirometry comparative study for toxicity determination has been performed. Seven organic and five inorganic toxic compounds have been assessed for comparing both methods. Microtox proved to have a higher sensitivity to toxicants but was less representative of effects on activated sludge compared to respirometry. For instance, assays accomplished with LAS, a biodegradable reference surfactant, showed a toxic effect by Microtox but good biodegradability and no toxicity in respirometry. This could be explained by the different nature of the biological material used, as Microtox utilises the seawater Vibrio fischeri, whereas respirometry uses the bacterial consortium in activated sludge. For the evaluation of the potential toxicity of a compound on a WWTP, the preferred biological material be used should be activated sludge itself. Results obtained with any other biological material would be just an approach to reality.     

A Daphnia magna feeding bioassay as a cost effective and ecological relevant sublethal toxicity test for Environmental Risk Assessment of toxic effluents

Environmental Risk Assessment of chemical products and effluents within EC countries require the use of cost effective standardized toxicity tests that in most cases are restricted to acute responses to high doses. Thus, subtle ecological effects are underestimated. Here we propose a short-term one day Daphnia magna feeding inhibition test as a cost effective and ecological relevant sublethal bioassay. The sensitivity and reliability of the proposed bioassay was tested in the laboratory against standardized bacteria, algae growth, D. magna and fish acute toxicity test by using 16 chemical mixture x water type combinations that included four different water types fortified with four complex chemical mixtures. Water types included ASTM hard water and three selected effluents diluted 1/10 in water to mimic worse field situations that many overexploited arid river ecosystems suffer during summer months when effluents are discharged into them with little dilution. The results obtained denoted a greater sensitivity of the proposed feeding bioassay in 51 out of 65 tests performed with an average sensitivity 50 fold greater than that of the standardized tests. The greater differences were obtained for mixtures that included narcotic chemicals and the lowest differences for those containing pesticides. Furthermore, feeding responses to the studied contaminant mixtures behaved differently to increasing TOC content than those based on bioluminescent bacteria and algae. Increasing TOC coming from sewage treated effluents decrease toxicity to the latter bioassays but increased those of D. magna feeding bioassays. These results empathize the need to include additional bioassays to monitor more accurately and realistically the toxicity of effluents or surface waters dominated by effluent discharges, a quite common situation in America and Mediterranean arid regions.     

Selecting a battery of bioassays for ecotoxicological characterization of wastes

This study was conducted in France within the context of waste classification (Hazardous Waste Council Directive 91/689/EEC), and focused on "ecotoxic" property (H14). In 1998, an experimental test strategy was developed to assess ecotoxicological properties of wastes using a battery of six standardized bioassays. This combined direct and indirect approaches integrating two solid-phase tests: emergence and growth inhibition of Lactuca sativa (14 days), mortality of Eisenia fetida (14 days) and four standardized tests performed on water extracts from wastes: growth inhibition of Pseudokirchneriella subcapitata (3 days), inhibition of mobility of Daphnia magna (48 h), inhibition of reproduction of Ceriodaphnia dubia (7 days), inhibition of light emission of Vibrio fischeri (30 min). This study aimed to set up preliminary conclusions on relevancy of this experimental test strategy, based on data obtained since 1998. Results were analyzed from the combined use of Hierarchical Cluster Analysis, Principal Component Analysis and Nonlinear Mapping. These multivariate analyses clearly showed that it was possible to reduce this number of tests without changing the typology of the wastes. A battery of bioassays including one solid phase test and two tests performed on water extracts (L. sativa, V. fischeri and C. dubia) was found as an optimal solution for characterizing the toxicity of the studied wastes. This optimal battery represents a good basis for determining the H14 property.     

上海城市污水厂污泥的农用处置方法研究

历时一年的调查显示，上海11家城市污水厂的污泥中含有大量的植物养分，但是在某些时期重金属含量会超过国家《农用污泥质量标准》，为此采用生物法和化学法对污泥中的重金属进行了沥滤试验。沥滤7d后的结果表明，采用生物法双用浓H2SO4酸化具更好的滤除效果；在生物法沥滤重金属的过程中还具有酸化法所没有的泥消化功能。     

The combined toxic effects of nonpolar narcotic chemicals to Pseudokirchneriella subcapitata

This paper presents the toxicity data of 10 nonpolar narcotic chemicals on Pseudokirchneriella subcapitata (green algae) assessed by a new algal toxicity testing technique conducted under air-tight environment. Based on DO production, median effective concentration (EC50) varies from 1.73 mg/L (1-octanol) to 8,040 mg/L (2-propanol). The endpoint of algal growth rate reveals similar sensitivity as that from DO production. Compared to literature data, Pseudokirchneriella subcapitata and Nitrosomonas are apparently more sensitive to nonpolar narcotics than other organisms such as minnow, daphnia, and Tetrahymena pyriformis. Furthermore, good correlations between toxic effects observed from Pseudokirchneriella subcapitata and other aquatic organisms were found. Hence, algal toxicity test can be considered as a surrogate test for estimating the toxicity of nonpolar chemicals to fathead minnow, Microtox, activated sludge, Daphina magna, and Tetrahymena pyriformis. The combined effects of 13 binary mixtures of nonpolar chemicals were investigated using both additive-index method and isobologram analysis. Overall speaking, the joint actions between these chemicals are strictly additive. Model analyses indicate that these compounds act on identical reaction sites or receptors, which verify that these chemicals are of the same toxicity mechanism (narcosis).     

复合硫杆菌对污泥重金属生物淋滤机制的影响

对污泥生物淋滤过程中各种重金属元素的形态变化情况进行了测定,借助重金属化学形态转化规律,间接判断污泥重金属生物淋滤的转化机制。试验结果显示,Cu、Ni、Cd的生物淋滤是以直接机制为主的,Zn的生物淋滤过程是以间接机制为主的,Cr、Pb的生物淋滤过程属于混合机制,但其混合机理是有差别的。     

Cadmium phytotoxicity: Quantitative sensitivity relationships between classical endpoints and antioxidative enzyme biomarkers

In this work, cadmium phytotoxicity and quantitative sensitivity relationships between different hierarchical endpoints in plants cultivated in a contaminated soil were studied. Thus, germination rate, biomass growth and antioxidative enzyme activity (i.e. superoxide dismutase, peroxidase, catalase and glutathione reductase) in three terrestrial plants (Avena sativa L., Brassica campestris L. cv. Chinensis, Lactuca sativa L. cv. hanson) were analyzed. Plant growth tests were carried out according to an International Standard Organization method and the results were analyzed by ANOVA followed by Williams' test. The concentration of Cd2+ that had the smallest observed significant negative effect (LOEC) on plant biomass was 6.25, 12.5 and 50 mg Cd/kg dry soil for lettuce, oat and Chinese cabbage, respectively. Activity of all enzymes studied increased significantly compared to enzyme activity in plant controls. For lettuce, LOEC values (mg Cd/kg dry soil) for enzymic activity ranged from 0.05 (glutathione reductase) to 0.39 (catalase). For oat, LOEC values (mg Cd/kg dry soil) ranged from 0.19 (for superoxide dismutase and glutathione reductase) to 0.39 (for catalase and peroxidase). For Chinese cabbage, LOEC values (mg Cd/kg dry soil) ranged from 0.19 (peroxidase, catalase and glutathione reductase) to 0.39 (superoxide dismutase). Classical (i.e. germination and biomass) and biochemical (i.e. enzyme activity) endpoints were compared to establish a sensitivity ranking, which was: enzyme activity>biomass>germination rate. For cadmium-soil contamination, the determination of quantitative sensitivity relationships (QSR) between classical and antioxidative enzyme biomarkers showed that the most sensitive plant species have, generally, the lowest QSR values.     

Cadmium assimilation in the terrestrial isopod, Porcellio dilatatus--is trophic transfer important?

Terrestrial isopods have become important tools for the ecotoxicological assessment of metal-contaminated soils. Their value as an invertebrate model is partly because of their extraordinary capacity to bioaccumulate toxic metals from the environment. Replication of this accumulation process in the laboratory has in the past relied on the amendment of organic food substrates through the addition of inorganic metal salts. However, the bioavailability of the metals when presented through doping regimes may differ from the bioavailability of metals in nature, because over time metals become biologically compartmentalised and form complexes with organic molecules. This study examines the differential bioavailability of Cd to the terrestrial isopod, Porcellio dilatatus, when presented as either a Cd-amended diet or pre-incorporated biologically into lettuce (Lactuca sativa). Isopods were either provided with lettuce contaminated superficially with Cd(NO(3))(2) or lettuce grown hydroponically in growth media containing 100 microM Cd(NO(3))(2). Assimilation efficiency of Cd was greater among isopods that were fed the amended diet (71%, S.E.=7%), than among isopods feeding on biologically contaminated lettuce (52%, S.E.=5%) and demonstrates that speciation of Cd is likely to influence the rate of Cd assimilation and accumulation in a laboratory test.     

Comparison of several microbiological toxicity screening tests

Four short-term microbiological toxicity screening tests were compared using the following test chemicals: 3,5 dichlorophenol, cetyl trimethyl ammonium chloride, sodium lauryl sulfate, phenol, copper(II) sulfate, mercury(II) chloride and zinc(II) sulfate. These seven chemicals represent a wide range of toxicity. The methods examined were Beckman's Microtox system, the Spirillum volutans motility test, inhibition of respiratory activity of activated sludge, and inhibition of activated sludge TTC-dehydrogenase activity.The results obtained indicate that each method has its own toxicity sensitivity pattern, and among the substances tested, only mercury(II) chloride and phenol were ranked equally by the four methods as the most and the least toxic chemical, respectively. In a tentative ranking of the methods, according to sensitivity, the Microtox test came out as the most sensitive test, followed by the Spirillum test, which in turn appeared more sensitive than the two sludge tests. This ranking has meaning only in a statistical sense, however, and the variable nature of the results support the philosophy that for assessing toxicity more thoroughly, a battery of several tests is required.     

Toxicity assessment of common xenobiotic compounds on municipal activated sludge: comparison between respirometry and Microtox

The toxicity of four xenobiotic compounds 3,5-dichlorophenol, formaldehyde, 4-nitrophenol and dichloromethane, representative of industrial wastewater contaminants was evaluated by a simple respirometric procedure set up on the basis of OECD Method 209 and by the Microtox bioassay. Very good reproducibility was observed for both methods, the variation coefficients being in the range of 2-10% for the respirometric procedure and 6-15% for Microtox, values that can be considered very good for a biological method. Comparison of EC(50) data obtained with the two methods shows that in both cases 3,5-dichlorophenol is more toxic than other compounds investigated and dichloromethane has a very low toxicity value. Intermediate EC(50) values were found for the two other chemicals, formaldehyde and 4-nitrophenol. Moreover, the Microtox EC(50) values are generally lower (except for dichloromethane) than the respirometric ones: these differences could be explained by the fact that the Microtox method uses a pure culture of marine species and, therefore, should not necessarily be expected to behave like a community of activated sludge bacteria. In conclusion, both methods can be usefully applied for toxicity detection in wastewater treatment plants but it is advisable to take into account that Microtox is more sensitive than respirometry in estimating the acute toxicity effect on the biomass operating in the plant.     

生物沥浸改性后污泥基生物炭对Pb^2+和Cd^2+的吸附

为探索利用生物沥浸改性后污泥基生物炭去除废水中Pb^2+和Cd^2+的可行性,以生物沥浸改性后污泥为原料制备生物炭,并测定了该材料的性能和吸附特性。通过单因素试验研究了pH值、生物炭投加量对吸附Pb^2+、Cd^2+性能的影响,并利用吸附动力学和热力学来揭示吸附机制。试验结果表明,经生物沥浸改性后污泥基生物炭的重金属含量降低;吸附Pb^2+和Cd^2+的最佳pH值分别为5、6,最佳生物炭投加量均为2 g/L;其对Pb^2+和Cd^2+的吸附均能较好地符合准二级动力学方程和Langmuir等温线模型,在25、35、45℃三种温度下对Pb^2+、Cd^2+的饱和吸附量分别为30.68、39.95、43.93 mg/g和19.82、28.58、32.29 mg/g,吸附过程均主要以物理吸附为主,且为吸热过程。     

Dewatering characteristics of sludge conditioned by surfactant with bioleaching pretreatment

In view of the merits of bioleaching and surfactant for sludge treatment, the possibility of surfactant with bioleaching pretreatment applied to improve sludge dewaterability was investigated in this work. The results showed that cetyl trimethyl ammonium chloride (CTAC) with bioleaching pretreatment was highly efficient in dewatering sludge. The optimal CTAC dosage and bioleaching pH for this combination were 120 mg/g (dry solids) and 3.55, respectively, under which the water content of filtrated cake was 68.94% and the specific resistance to filtration was 0.12 × 10¹³ m/kg with a reduction of 94.92%. Although CTAC and bioleaching both had positive effects on sludge dewatering, their combination was more efficient. The significant enhancement of sludge dewaterability was mainly contributed from the breakage and charge neutralization of sludge flocs, and the release of extracellular polymeric substances (EPS). In addition, the positive correlation between supernatant EPS concentration and sludge dewaterability was observed under experimental conditions.     

Effects of sewage sludge on pH and plant availability of metals in oxidising sulphide mine tailings

A field study was conducted adjacent to the tailings deposit of the Aitik copper mine in the north of Sweden to investigate the effects of sewage sludge on pH and plant availability of Al, Mn, Cu, Zn, Ni, Pb, As, Cr and Cd in the oxidising sulphide tailings. One treatment was supplied with sewage sludge, while the control received NPK-fertiliser. The tailings samples were collected at the beginning and the end of the growing season and extracted by NH(4)NO(3), NH(4)Ac-EDTA and HNO(3). Plant tissue concentrations of the elements were determined in the above-ground parts of barley (Hordeum vulgare) and red fescue (Festuca rubra). The application of sewage sludge resulted in higher crop yields compared to the control, although the buffering capacity and the metal immobilising effect of the sludge were limited. The pH decreased from 6.6 to 4.3 in the control and from 6.4 to 4.8 in the sludge-treated tailings during the growing season, probably due to sulphide oxidation in the tailings. This resulted in increased levels of soluble elements in all treatments studied. Application of sewage sludge resulted in elevated levels of soluble Zn and lower values of soluble As and Cd in the unaltered tailings but increased levels of specifically adsorbed Cu, Ni and As in the oxidised tailings. This was partly reflected in the plants, as the application of sewage sludge resulted in 67 mg Zn kg(-1) in barley grains and 60 mg Zn kg(-1) in red fescue shoots, both values twice as high as the corresponding values in the control, but lower As contents in both straw (0.3 mg kg(-1)) and grain (0.06 mg kg(-1)) of barley compared to the control (0.6 and 0.2 mg kg(-1), respectively). In addition, red fescue grown in sludge-treated plots contained significantly higher levels of Al, Cu, Pb, As and Cr compared to the control. The levels of several metals in barley and red fescue grown in both treatments exceeded background values found in the literature. The Cu content in barley straw exceeded 100 mg kg(-1) in both treatments and might be toxic to grazing animals. Thus, this study suggests that adding sewage sludge to the mine tailings at Aitik would not counteract the effects of the sulphide oxidation in the tailings. Furthermore, using a sequential extraction technique proved preferable to using total metal analysis in order to predict plant uptake of the elements in the tailings.     

Effect of cigarette smoke on seed germination

The effect of cigarette smoke was studied on the germination of radish, kale, lettuce, amaranth, wheat, rice, barley and rye seeds. It was found that such smoke markedly retarded, in all cases, the rate of germination. Furthermore, cigarette smoke caused a retardation of the levels of certain enzymes (alpha-amylase or lysozyme) known to be significant in the germination of these seeds.     

Ecotoxicological tests assessment of soils polluted by chromium (VI) or pentachlorophenol

Carbon mineralisation and plant germination and growth (Lactuca sativa seeds) tests have been performed in two soils of different properties, experimentally spiked with pentachlorophenol (PCP) or Cr (VI), in concentrations between 0.001 and 1000 mg kg(-1). The evaluation has been done considering the following parameters of carbon transformation test: soil cumulative basal respiration after 14 days incubation, substrate induced respiration after 12 h of glucose addition, and, in the plant germination and growth test: number of germinated seeds, root elongation and total biomass produced. The most sensitive assay found in our work has been carbon mineralisation test, from which the lowest toxic concentrations were obtained (especially substrate induced respiration test). In the plant germination and growth test, the measurement of root elongation has shown the best sensitiveness, followed by plant biomass and seed germination numbers. Regarding the contaminants, the highest toxicity, considering the minimum concentration with toxic effect, has been found in PCP (0.01 mg kg(-1)) in C mineralisation test in the granitic soil. For Cr, the minimum concentration with toxic effect has been 0.1 mg kg(-1), also in the C mineralisation test and the granitic soil. The granitic soil has shown more vulnerability to the pollutants assayed in the respiration test, whereas the calcareous soil has shown more vulnerability in the plant germination and growth test.     

A comparison of microbial bioassays for the detection of aquatic toxicants

The toxicity of 35 test chemicals was analyzed using two microbial bioassay systems. The commercially available Microtox Toxicity Analyzer System™ and the two-organism procedure of Tchan were used to determine the concentration of test chemicals resulting in a 50% reduction in response (EC₅₀). Both of the tests employed a luminescent bacterium while the procedure of Tchan also utilized an alga. Results from the two microbial tests were compared with available data obtained with fish toxicity bioassays and with each other. The Microtox™ procedure was somewhat more sensitive than the Tchan bioassay in detecting most of the test chemicals and fish bioassays were generally more sensitive than either of the microbial tests. As a notable exception, photosynthesis-inhibiting herbicides were detected at remarkably lower concentrations with the procedure of Tchan than any of the other bioassays. Potential applications for these tests are discussed.     

A simple system to rapidly monitor activated sludge health and performance

A set of four assays designed to rapidly measure the health and biodegradative performance of pulp and paper mill activated sludges was developed. Three of the assays are specific oxygen uptake rates (SOURs) that measure the normal "working" aeration tank BOD (biochemical oxygen demand) removal rate (SOURAT), a near-maximum BOD removal rate (SOURNMAX), and a rate (SOURTOX) used in combination with the SOURNMAX to indicate the presence of toxic or inhibitory substances. The fourth assay is the specific adenosine triphosphate (SATP) content of the sludge, used as a measure of its viable cell content. Fresh biomass (sludge) samples from one laboratory reactor and four mill biotreatment systems were fed raw mill effluents and used to evaluate the four-assay set. The SOURAT values of all systems were 10-40% of their SOURNMAX values: thus the SOURAT:SOURNMAX ratios indicate that each system's free biodegradative capacity was far greater than its operating rate. It was demonstrated using phenol that the SOURNMAX:SOURTOX ratio can indicate the presence of substances toxic or inhibitory to the biomass. The results also indicated that the SOURNMAX is a much better indicator of improving or worsening sludge performance and capacity than the SOURAT. SATP was shown to be a useful monitor of the proportion of viable cells in an activated sludge and a toxicity indicator complementary to the SOURNMAX:SOURTOX ratio and similar in principle to the commercial Microtox toxicity test. This four-assay set was also applied to three practical situations: (a) at-mill monitoring of a biotreatment system; (b) effects of cold storage on biomass; and (c) effects of decreased BOD loading on biomass.     

Environmental assessment of anaerobically digested sludge reuse in agriculture: Potential impacts of emerging micropollutants

Agricultural application of sewage sludge has been emotionally discussed in the last decades, because the latter contains organic micropollutants with unknown fate and risk potential. In this work, the reuse of anaerobically digested sludge in agriculture is evaluated from an environmental point of view by using Life Cycle Assessment methodology. More specifically, the potential impacts of emerging micropollutants, such as pharmaceuticals and personal care products, present in the sludge have been quantified. Four scenarios were considered according to the temperature of the anaerobic digestion (mesophilic or thermophilic) and the sludge retention time (20 or 10 d), and they have been compared with the non-treated sludge.From an environmental point of view, the disposal of undigested sludge is not the most suitable alternative, except for global warming due to the dominance (65-85%) of the indirect emissions associated to the electricity use. Nutrient-related direct emissions dominate the eutrophication category impact in all the scenarios (>71.4%), although a beneficial impact related to the avoidance of industrial fertilisers production is also quantified (up to 6.7%). In terms of human and terrestrial toxicity, the direct emissions of heavy metals to soil dominate these two impact categories (>70%), and the contribution of other micropollutants is minimal. Moreover, only six (Galaxolide, Tonalide, Diazepam, Ibuprofen, Sulfamethoxazole and 17α-ethinyloestradiol) out of the 13 substances considered are really significant since they account for more than 95% of the overall micropollutants impact.     

Toxicity assays: a way for evaluating AOPs efficiency

The technical feasibility and performance of photocatalytic degradation of aqueous methomyl (50 mg/L) have been studied at pilot scale in two well-defined systems of special interest because natural-solar UV light can be used: heterogeneous photocatalysis with titanium dioxide and homogeneous photocatalysis by photo-Fenton. The pilot plant is made up of compound parabolic collectors specially designed for solar photocatalytic applications. Experimental conditions allowed pesticide disappearance, degree of mineralisation and toxicity achieved in the two photocatalytic systems to be compared. Total disappearance of methomyl is attained by photo-Fenton in 60 min and by TiO2 in 100 min. Hundred percent of nitrogen and sulphur are recovered as ammonium and sulphate. By contrast, complete mineralisation of total organic carbon (TOC) is not achieved even after quite a long time (more than 300 min). Three different bioassays (Vibrio fischeri, Daphnia magna and a Microalga) have been used for testing the progress of toxicity during treatment. All remained toxic down to very low-pesticide concentrations and in some bioassays were still toxic after total disappearance of the pesticide. Only if treatment is maintained throughout enough mineralisation (i.e. TOC disappearance), the toxicity is reduced to below the threshold (EC50%).