Acute and sublethal toxicity tests to monitor the impact of leachate on an aquatic environment

In this study, a specific landfill leachate (1200 mg l(-1) COD and 600 mg l(-1) BOD(5)) was used to develop a standardised short-term acute and longer-term sublethal ex-situ toxicity testing programme, in order to determine the potential ecological implications of leaching contaminants reaching the water table. Bioassays were undertaken with juvenile Gammarus pulex and Asellus aquaticus macro-invertebrates. Preliminary acute test variables included static and static renewed flow rates for 96-h, starved and fed specimens, and aerobic and oxygen depleting conditions. However, regardless of any test variable, the lethal concentration (LC(50)) for A. aquaticus remained at 12.3% v/v leachate in deionized water, whilst that for G. pulex was only 1%. Sublethal toxicity was judged on the basis of frequency of births and the growth rate of newly born individuals. Tests showed that even a dilution as high as 1:66- would influence the fecundity of a Gammarus population, whilst a dilution of 1:20 would affect the size of an Asellus breeding colony.     

Toxicities and tolerances of Cd, Cu, Pb and Zn in a primary producer (Isochrysis galbana) and in a primary consumer (Perna viridis)

Studies on toxicities and tolerances of cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) in the brown alga Isochrysis galbana and in the green-lipped mussel Perna viridis were conducted by short-term bioassays using endpoints growth production and mortality, respectively. The 5-day EC(50) and 24-h LC(50) of these heavy metals were determined in the brown alga and mussel, respectively. The EC(50) values calculated for the alga were 0.74 mg/l for Cd, 0.91 mg/l for Cu, 1.40 mg/l for Pb and 0.60 mg/l for Zn. The LC(50) values for the mussels were 1.53 mg/l for Cd, 0.25 mg/l for Cu, 4.12 mg/l for Pb and 3.20 mg/l for Zn. These LC(50) values were within the concentration ranges as reported by other authors who used P. viridis as the test organism. Based on these EC(50) and LC(50) values, the alga was most sensitive to Zn, followed by Cd, Cu and Pb while the mussel was most sensitive to Cu, followed by Cd, Zn and Pb. Differences in the trophic levels, metal handling strategies, biology and ecology of the primary producer (brown alga) and the primary consumer (mussel) are believed to be the plausible causes for the different toxicities and tolerances of the metals studied.     

Comparative toxicity of solid waste leachates to Daphnia magna

The comparative acute and chronic toxicities of municipal, industrial, coal-fired power plant, and synthetic fuel solid waste leachates to the aquatic invertebrate Daphnia magna were determined. Seventeen leachates were laboratory-derived by extraction with 0.5 N acetic acid, and one, an arsenic-contaminated groundwater, was naturally derived. The acute toxicity 48-h LC₅₀ estimates (concentrations of the test materials that killed 50% of the test organisms in 48 h) ranged from 0.0005% for a dye waste leachate to > 100% (i.e., undiluted) for a coal gasification waste leachate. The most toxic leachates were derived from a dye waste, a plater's waste, and the arsenic-contaminated groundwater, and the least toxic were from a soybean process cake waste and a coal gasification waste. Of the 16 leachates used in the 28 day chronic toxicity tests, only the arsenic-contaminated groundwater and a municipal sewage sludge significantly affected D. magna reproduction. The former material at 1.0% concentration and the sludge at 0.1% caused about an 88% reduction in the numbers of young produced by exposed females compared with controls. Generally, the industrial waste leachates were more toxic than those from the power plant and synthetic fuel wastes. Because its presence in the test materials interfered with interpretation of the results, acetic acid may not be an appropriate extraction medium for preparation of leachates.     

Toxicokinetics/toxicodynamics of arsenic for farmed juvenile milkfish Chanos chanos and human consumption risk in BFD-endemic area of Taiwan

This paper presents a toxicokinetic/toxicodynamic analysis to appraise arsenic (As) bioaccumulation in farmed juvenile milkfish Chanos chanos at blackfoot disease (BFD)-endemic area in Taiwan, whereas probabilistic incremental lifetime cancer risk (ILCR) and hazard quotient (HQ) models are also employed to assess the range of exposures for the fishers and non-fishers who eat the contaminated fish. We conducted a 7-day exposure experiment to obtain toxicokinetic parameters, whereas a simple critical body burden toxicity model was verified with LC50(t) data obtained from a 7-day acute toxicity bioassay. Acute toxicity bioassay indicates that 96-h LC50 for juvenile milkfish exposed to As is 7.29 (95% CI: 3.10-10.47) mg l(-1). Our risk analysis for milkfish reared in BFD-endemic area indicates a low likelihood that survival is being affected by waterborne As. Human risk analysis demonstrates that 90%-tile probability exposure ILCRs for fishers in BFD-endemic area have orders of magnitude of 10(-3), indicating a high potential carcinogenic risk, whereas there is no significant cancer risk for non-fishers (ILCRs around 10(-5)). All predicted 90%-tiles of HQ are less than 1 for non-fishers, yet larger than 10 for fishers which indicate larger contributions from farmed milkfish consumptions. Sensitivity analysis indicates that to increase the accuracy of the results, efforts should focus on a better definition of probability distributions for milkfish daily consumption rate and As level in milkfish. Here we show that theoretical human health risks for consuming As-contaminated milkfish in the BFD-endemic area are alarming under a conservative condition based on a probabilistic risk assessment model.     

Toxicity of copper in sewage sludge

Sewage sludge is a source of organic matter and nutrients, but a major obstacle for its recycling is that the municipal wastewater sludge has low but significant levels of contaminants. This investigation, on the acute toxicity of copper in sewage sludge, was conducted with three organisms, Daphnia magna, Lemna minor and Raphanus sativus (seeds). The toxicity of the leakage water from sewage sludge spiked with CuSO(4) was studied for 64 days. The toxicity increased during the first 8-16 days and then started to decrease. The first increase in toxicity was due to ammonia, but after 32 days, a dose-related effect of copper was found. After 64 days, L. minor had an EC50 of 3800 mg Cu/kg dw for 7 days growth inhibition, a LOEC of 3200 mg Cu/kg dw and a NOEC of 1600 mg Cu/kg dw. D. magna had an EC50 of 18100 mg Cu/kg dw (24-h immobility) and a NOEC of 12800 mg Cu/kg dw. Root elongation of R. sativus was reduced at 25600 mg Cu/kg dw. Both for Daphnia and Lemna, the pH of the leakage water had an effect of the toxicity. This means that chemical speciation and bioavailability is very important for the hazard assessment of copper in sludge and soil.     

Effects of cadmium and zinc on larval growth and survival in the ground beetle,Pterostichus oblongopunctatus

Carabid beetles, like Pterostichus oblongopunctatus, living in metal contaminated areas may be exposed to elevated levels of metals within their diets. However, when compared to other second order consumers, they have one of the lowest observed levels of metals, indicating methods of detoxification to deal with such toxicants. In this study, we investigated if chronic, multigenerational exposure to metals leads to resistance to toxic metal concentrations, and if so, what are the costs associated with them. Adult organisms were collected from two sites, a polluted and a reference site near Olkusz, in southern Poland. These adults were immediately mated, and eggs were collected twice weekly to assess the effects in the larvae of the F(1) generation. Larvae were randomly exposed to one of four artificial mediums: control, 50 mg kg(-1) Cd, 500 mg kg(-1) Zn, and a combined treatment of 50 mg kg(-1) Cd and 500 mg kg(-1) Zn to investigate possible interactions. Individuals were sacrificed at 10, 30, and 40 days. Although metals were not accumulated in larvae (p>0.05), larvae fed the Cd or the Zn treatment grew significantly slower, and had the lowest survival rate (p<0.05) in respect to control. Out of metal treated animals, those on the combined treatment of CdZn grew the quickest and had the highest observed survival (p<0.05). Although previous studies have demonstrated changes in adult population parameters under chronic, multigeneration exposure to toxic metal concentrations, our study did not reveal any changes in the larval stage.     

Inhibition of microbial activity of activated sludge by ammonia in leachate

Leachate samples were collected from the West New Territory Landfill (WENT), Hong Kong, and characterized in the laboratory. The analytical results confirmed that it has a typical nature of aged leachate with a low BOD₅/COD ratio of 0.22 and a high strength of ammonia-nitrogen around 5 g/L. A lab-scale study was conducted to investigate the inhibition of microbial activity of the activated sludge. In the first test, glucose-based synthetic wastewater was used in two parallel reactors. The experimental results demonstrated that COD removal declined from 95.1 to 79.1% and the dehydrogenase activity of the sludge decreased from 11.04 to 4.22 μg TF/mg mixed liquor suspended solids (MLSS), when the ammonia-nitrogen concentration increased from 50 mg/L to 800 mg/L progressively. The remaining NH₃⁺-N residue in the treated wastewater increased from 0.58 mg/L to 649 mg/L extensively. In the second test, mixed wastewater samples containing glucose and raw leachate were fed into six parallel biological reactors and operated on batch mode. The experimental results showed COD removal decreased from 97.7 to 78.1% and the dehydrogenase activity decreased from 9.29 to 4.93 μg triphenyl formazon (TF)/mg MLSS, respectively, when the ammonia-nitrogen concentration increased within the same range. Microbial inhibition could also be substantiated by a decrease of specific oxygen uptake rate (SOUR) from 68 to 45 mg O₂/g MLSS. These results suggested leachate containing high-strength ammonia-nitrogen should be pretreated to an acceptable NH₄⁺-N level before it is fed into biological reactors.     

Aerobic degradation of 2,4,6-TCP content in ECF bleached effluent

Elemental chlorine-free (ECF) bleach effluents from kraft mill are characterised by: a chemical organic demand/biological organic demand (COD/BOD(5)) ratio of 4, chlorophenol content with low chlorine substitution, and toxicity. The effect of increasing the concentration of 2,4,6-trichlorophenol (2,4,6-TCP) content in ECF bleaching sequence effluent on the degradative activity of bacterial communities present in an aerobic system treatment was studied. An aerobic lagoon (AL) was used as a typical secondary treatment of kraft-mill effluent. AL displays a high performance of BOD(5) degradation (up to 90%); however, only 40% of the COD was removed. Simultaneously, the AL system shows a high ability to biodegrade 2,4,6-TCP up to 237 mg/l day. Kinetic parameters of the 2,4,6-TCP biodegradation by aerobic bacteria were determined. The K(s) and K(i) values were 34.3 and 50 mg/l 2,4,6-TCP, respectively. Moreover, the tolerance of aerobic bacteria was observed up to 1.3 g/l 2,4,6-TCP.     

Toxicological responses in Laeonereis acuta (annelida, polychaeta) after arsenic exposure

Several environmental pollutants, including metals, can induce oxidative stress. So, the objective of this study was to evaluate the effects of arsenic (As(III), as As(2)O(3)) on the antioxidant responses in the polychaete Laeonereis acuta. Worms were exposed to two environmentally relevant concentrations of As, including the highest previously allowed by Brazilian legislation (50 microg As/l). A control group was kept in saline water (10 per thousand) without added metal. It was observed that: (1) a peak concentration of lipid peroxide was registered after 2 days of exposure to 50 microg As/l (61+/-3.2 nmol CHP/g wet weight) compared to the control group (43+/-4.5 nmol CHP/g wet weight), together with a lowering of the activity of the antioxidant enzyme catalase (-47 and -48%, at 50 or 500 microg As/l respectively) and a higher superoxide dismutase activity (+305% at 50 microg As/l with respect to the control group); (2) a lower conjugation capacity through glutathione-S-transferase activity was observed after 7 days of exposure to 50 microg As/l (-48% compared to the control group); (3) a significant increase in As concentration was verified after 1 week of exposure to both As concentrations (50 and 500 microg/l); (4) worms exposed to As showed a limited accumulation of related methylated As species and the levels of non-toxic As species like arsenobetaine (AsB) and arsenocholine (AsC) remained unchanged during the exposure period when compared with the controls. Overall, it can be concluded that As interfered in the antioxidant defense system of L. acuta, even at low concentrations (50 microg/l) that Brazilian legislation previously considered safe. The fact that worms exposed to As showed high levels of methylated As species indicates the methylation capability of L. acuta, although the high levels of inorganic As suggest that not all the administered As(III) (as As(2)O(3)) is completely removed or biotransformed after 7 days of exposure.     

Contribution of (222)Rn-bearing water to indoor radon and indoor air quality assessment in hot spring hotels of Guangdong, China

This study investigates the contribution of radon (²²²Rn)-bearing water to indoor ²²²Rn in thermal baths. The ²²²Rn concentrations in air were monitored in the bathroom and the bedroom. Particulate matter (PM, both PM₁₀ and PM_2.5) and carbon dioxide (CO₂) were also monitored with portable analyzers. The bathrooms were supplied with hot spring water containing 66-260 kBq m⁻³ of ²²²Rn. The results show that the spray of hot spring water from the bath spouts is the dominant mechanism by which ²²²Rn is released into the air of the bathroom, and then it diffuses into the bedroom. Average ²²²Rn level was 110-410% higher in the bedrooms and 510-1200% higher in the bathrooms compared to the corresponding average levels when there was no use of hot spring water. The indoor ²²²Rn levels were influenced by the ²²²Rn concentrations in the hot spring water and the bathing times. The average ²²²Rn transfer coefficients from water to air were 6.2 × 10⁻⁴-4.1 × 10⁻³. The 24-h average levels of CO₂ and PM₁₀ in the hotel rooms were 89% and 22% higher than the present Indoor Air Quality (IAQ) standard of China. The main particle pollutant in the hotel rooms was PM_2.5. Radon and PM₁₀ levels in some hotel rooms were at much higher concentrations than guideline levels, and thus the potential health risks to tourists and especially to the hotel workers should be of great concern, and measures should be taken to lower inhalation exposure to these air pollutants.     

Comparative Sensitivity to the Insecticides Deltamethrin and Esfenvalerate of Some Aquatic Insect Larvae (Ephemeroptera and Odonata) and Daphnia magna

Sensitivity to the pyrethroids deltamethrin and esfenvalerate (aqueous solution) and LC₅₀ have been determined in acute (96-h) toxicological tests on mayfly larvae (Cloeon dipterum and Caenis miliaria), damselfly larvae (Lestes sponsa and Cordulia aenea), and juveniles from a laboratory culture of Daphnia magna. Sensitivity to deltamethrin increases in the series C. aenea (Odonata) < D. magna (Cladocera) < L. sponsa (Odonata) < C. miliaria(Ephemeroptera) < C. dipterum (Ephemeroptera), and that to esfenvalerate, in the series C. aenea < D. magna < L. sponsa C. miliaria C. dipterum. The values of LC₅₀ about 0.01 g/l determined for mayfly larvae are below those known for various hydrobionts from the literature, indicating a very high sensitivity of these insects to pyrethroids.     

Reproduction of the Siberian spruce in the timberline ecotone of the Iremel’ Massif

Quantitative and qualitative characteristics of cones and seeds and the emergence and survival of seedlings in the Siberian spruce (Picea obovata Ledeb.) were studied in different parts of the timberline ecotone on the Iremel’ Massif, the Southern Urals, from 2003 to 2006. In the productive year 2003, a decrease in the number of cones (from 110000 to 3000 per hectare), the yield of seeds (from 13 × 10⁶ to 0.09 × 10⁶ per hectare), and their laboratory germination rate (from 34 to 14%) with an increase in elevation above sea level was recorded. In 2004, significant differences in the number of seedlings per hectare were revealed between the lower, middle, and upper parts of the ecotone (85 200, 19 800, and 0–400 ind., respectively). It has been shown that the greater part of seedlings (76–96%) perish during the first cold period. New generations of spruce trees are formed only after productive years. Seed production and seedling mortality in spruce apparently depend not only on air temperature and soil moistening but also on snow depth and soil temperature, which change more abruptly along the altitudinal gradient.     

澎溪河回水区水体叶绿素a含量与流速相关关系研究

以三峡库区支流澎溪河回水区为特征区域,研究三峡水库回水区水动力条件对水体富营养化的影响。根据三峡水库的调蓄过程,在175 m高水位、小流量、低污染负荷和145 m低水位、大流量、高污染负荷两种设计条件下模拟澎溪河水体断面平均高锰酸盐指数（CODMn）、总氮（TN）、总磷（TP）浓度和叶绿素a（Chl a）含量。研究表明：澎溪河回水段从上游到下游的CODMn、TN、TP浓度有增加的趋势,在接近河口处有趋同于长江水污染物浓度的趋势。145 m低水位的大流量、高污染负荷设计条件下Chl a含量较高,在弯道下游和河道水面开阔的高阳、双江大桥断面的流速减缓区段,出现Chl a含量峰值。拟合澎溪河水体流速及Chl a含量相关关系,在枯丰两种设计条件下建立水体流速(V)与Chl a含量(S)的相关关系式,分别为S=0186 0〖DK〗·V-0353 6(相关系数r2=0838 0)和S=10844 9〖DK〗·V-0293 9(相关系数r2=0825 4)。模拟结果表明：加大水体流速有利于抑制Chl a含量。通过改变水动力条件控制水体富营养化具有可行性     

Chemical evaluation of potable water in Eastern Qinghai Province, China: human health aspects

Electric conductivity, pH, COD(Mn), nutrient concentration, chloride and sulfate concentrations, total dissolved sodium, potassium, calcium, magnesium, manganese, iron, cadmium, copper, arsenic, nickel, zinc, chromium and lead were evaluated to clarify concerns about the quality and safety of water used for drinking purposes in Qinghai Province, China. For this purpose, 12 water samples were collected from different villages, Qinghai (Koko Nor) Lake and medicinal springs close to the Town of Pingang during a study visit to China in 2003. The results showed that National Chinese and WHO drinking water standards were exceeded for nutrient concentration (3.2 mg l(-1) of TOT-N and 0.2 mg l(-1) of TOT-P) from Qinghai Lake. The presence of elevated electric conductivity (550 mS m(-1)) in mineral water resort samples should be a matter of a public concern. Also, samples from medicinal springs showed high concentrations of Fe (up to 1.9 mg l(-1)), As (up to 0.1 mg l(-1)) and Ni (0.05 mg l(-1)), which may be detrimental for human health if the water is consumed on a daily basis. The concentrations of Cu, Cd, Cr and Pb did not exceed the National Chinese and WHO drinking water standards, and therefore, water from the sampling area does not pose any significant threat to the consumers' health regarding these metals.     

A quantitative estimate of nonsmokers' lung cancer risk from passive smoking

This work presents a quantitative assessment of nonsmokers' risk of lung cancer from passive smoking. The estimates given should be viewed as preliminary and subject to change as improved research becomes available. It is estimated that U.S. nonsmokers are exposed to from 0 to 14 mg of tobacco tar per day, and that the typical nonsmoker is exposed to 1.4 mg per day. A phenomenological exposure-response relationship is derived, yielding 5 lung cancer deaths per year per 100,000 persons exposed, per mg daily tar exposure. This relationship yields lung cancer mortality rates and mortality ratios for a U.S. cohort which are consistent to within 5% with the results of both of the large prospective epidemiological studies of passive smoking and lung cancer in the United States and Japan. Aggregate exposure to ambient tobacco smoke is estimated to produce about 5000 lung cancer deaths per year in U.S. nonsmokers aged ≥ 35 yr, with an average loss of life expectancy of 17 ± 9 yr per fatality. The estimated risk to the most-exposed passive smokers appears to be comparable to that from pipe and cigar smoking. Mortality from passive smoking is estimated to be about two orders of magnitude higher than that estimated for carcinogens currently regulated as hazardous air pollutants under the federal Clean Air Act.     

The toxicity of copper contaminated soil using a gnotobiotic Soil Multi-species Test System (SMS)

A gnotobiotic multi-species study was designed to consist of a food-web of soil-dwelling animals. The food-web was exposed to five concentrations copper (Cu) spiked soil for three exposure durations i.e. 28, 56 and 84 days. Based on multivariate analysis the food-web was significantly affected by Cu exposure at and above 300 mg Cu kg(-1) soil (lowest tested concentration). The number of animals present in the 2500 mg Cu kg(-1) (highest tested concentration) was at all sampling occasions below the starting point level. Based on analysis of the individual species the lowest 10% effect concentration (EC10) observed was 50 mg Cu kg(-1) soil, for Enchytraeus crypticus. Using the EC10 for the individual species the HC5 (Hazard Concentration at the 5% level) was estimated to be between 25 and 36 mg Cu kg(-1) soil, depending on the exposure duration. A similar experiment but using a reduced design was performed employing soil contaminated with Cu in the field more than 80 years ago. The trend in the field-contaminated soil was similar to that observed for the spiked soil.     

BOD-DO modeling and water quality analysis of a waste water outfall off Kochi, west coast of India

Water quality scenarios around an offshore outfall off Kochi were simulated using MIKE21 water quality model, assuming a high Biochemical Oxygen Demand (BOD=50 mg l(-1)) effluent discharge. The discharge is introduced into the model through an outfall located at a distance of 6.8 km from the shore at a depth of 10 m. Three scenarios were simulated with different discharge rates such as 2, 5 and 10 m3 s(-1), with BOD load of 8640, 21,600 and 43,200 kg day(-1) respectively. Model simulations were carried out to estimate the assimilation capacity of the waters off Kochi for the three discharge rates. The results show that for 10 m3 s(-1) effluent discharge, the initial BOD of 50 mg l(-1) reduced to 3.33 mg l(-1) at the outfall after 48 h. High BOD values were confined to an elliptical area of approximately 8 km2 around the outfall. Based on this, the assimilative capacity of the waters off Kochi in terms of BOD can be estimated as 38,000 kg day(-1). It is suggested that offshore waters could be used as a feasible alternative to the Kochi backwaters for the disposal of treated effluent.     

Fate and effect of 2,4-dichlorophenol on biological treatment processes

The fate and effect of continuous feeding of increasing doses of 2,4-dichlorophenol (DCP) were investigated. At the beginning of each run, a drop in the efficiency of the system was recorded, after which a steady state was obtained. Continuous feeding of DCP up to a concentration of 50 mg L^?1 reduced BOD₅ and COD removal rates by 27% and 29%, respectively. Application of shock doses of DCP showed that adaptation increases the capacity of biological systems for removing toxic chemicals.     

Characteristics of PM10, PM2.5, CO2 and CO monitored in interiors and platforms of subway train in Seoul, Korea

This study was performed to investigate the concentration of PM₁₀ and PM_2.5 inside trains and platforms on subway lines 1, 2, 4 and 5 in Seoul, KOREA. PM₁₀, PM_2.5, carbon dioxide (CO₂) and carbon monoxide (CO) were monitored using real-time monitoring instruments in the afternoons (between 13:00 and 16:00). The concentrations of PM₁₀ and PM_2.5 inside trains were significantly higher than those measured on platforms and in ambient air reported by the Korea Ministry of Environment (Korea MOE). This study found that PM₁₀ levels inside subway lines 1, 2 and 4 exceeded the Korea indoor air quality (Korea IAQ) standard of 150 μg/m³. The average percentage that exceeded the PM₁₀ standard was 83.3% on line 1, 37.9% on line 2 and 63.1% on line 4, respectively. PM_2.5 concentration ranged from 77.7 μg/m³ to 158.2 μg/m³, which were found to be much higher than the ambient air PM_2.5 standard promulgated by United States Environmental Protection Agency (US-EPA) (24 h arithmetic mean: 65 μg/m³). The reason for interior PM₁₀ and PM_2.5 being higher than those on platforms is due to subway trains in Korea not having mechanical ventilation systems to supply fresh air inside the train. This assumption was supported by the CO₂ concentration results monitored in tube of subway that ranged from 1153 ppm to 3377 ppm. The percentage of PM_2.5 in PM₁₀ was 86.2% on platforms, 81.7% inside trains, 80.2% underground and 90.2% at ground track. These results indicated that fine particles (PM_2.5) accounted for most of PM₁₀ and polluted subway air. GLM statistical analysis indicated that two factors related to monitoring locations (underground and ground or inside trains and on platforms) significantly influence PM₁₀ (p < 0.001, R² = 0.230) and PM_2.5 concentrations (p < 0.001, R² = 0.172). Correlation analysis indicated that PM₁₀, PM_2.5, CO₂ and CO were significantly correlated at p < 0.01 although correlation coefficients were different. The highest coefficient was 0.884 for the relationship between PM₁₀ and PM_2.5.     

Identification of metal toxicity in sewage sludge leachate

Sewage sludge is a source of organic matter and nutrients with the potential for being used as a fertilizer. However, metals in sewage sludge might accumulate in soil after repeated sludge applications, and metal concentrations might reach concentrations that are toxic to microorganisms, soil organisms and/or plants. This toxicity might change with time due to kinetic factors or abiotic factors such as freezing, drying or rainfall. The objective of this study was to determine toxicity of sewage sludge leachate from a lysimeter with 50 cm of sludge applied. Attempts were also made to identify the cause of toxicity of the sludge leachate by toxicity identification and evaluation (TIE) techniques. Sludge leachate was collected monthly during 1 experimental year (August 2001 to August 2002). Metal concentrations were analysed, and the toxicity was determined with Daphnia magna (48-h immobility). The effect of EDTA or sodium thiosulphate addition, filtration through a CM-resin or a Millex-resin on toxicity was also tested. The results showed that toxicity of the sludge leachate apparently varied during the year, and that filtration through the CM-resin reduced most of the toxicity followed by the addition of EDTA. None of the other treatments reduced the toxicity of the sludge leachate. This indicated that one or more metals were responsible for the observed toxicity. Further calculations of toxic units (TU) suggested that Zn contributed most to the toxicity. Results also indicated that Ca concentrations in the sludge leachate reduced the toxicity of Zn.