Right ventriculoarterial coupling surrogates and long-term survival in LVAD recipients: Results of the ASSIST-ICD multicentric registry.

BACKGROUND: Prediction of outcomes remains an unmet need in LVAD candidates. Development of right heart failure portends an excess in mortality but imaging parameters of right ventricular systolic function have failed to demonstrate a prognostic role. By integrating pulmonary pressure, right ventriculoarterial coupling could fill this gap. METHODS: The ASSIST-ICD registry was used to test right ventriculoarterial coupling surrogate parameters at implantation for the prediction of all-cause mortality. RESULTS: The ratio of the tricuspid annular plane systolic excursion over the estimated systolic pulmonary pressure (TAPSE/sPAP) was not associated with long-term survival in univariate analysis (p = 0.89), neither was the pulmonary artery pulsatility index (PAPi) (p = 0.13). Conversely, the ratio of the right atrial pressure over the pulmonary capillary wedge pressure (RAP/PCWP) was associated with all-cause mortality (p <0.01). After taking tricuspid regurgitation severity, LVAD indication, LVAD model, age, blood urea nitrogen, and pulmonary vascular resistance into account, RAP/PCWP remained associated with survival (HR 1.35 [1.10 - 1.65], p <0.01). CONCLUSION: Among pre-implant RVAC surrogates, only RAP/PCWP was associated with long-term all-cause mortality in LVAD recipients. This association was independent of established risk factors.

Massive increase, spread, and exchange of extended spectrum ß-lactamase-encoding genes among intestinal Enterobacteriaceae in hospitalized children with severe acute malnutrition in Niger.

BACKGROUND: From the time of CTX-M emergence, extended-spectrum ß-lactamase-producing enterobacteria (ESBL-E) have spread worldwide in community settings as well as in hospitals, particularly in developing countries. Although their dissemination appears linked to Escherichia coli intestinal carriage, precise paths of this dynamic are largely unknown. METHODS: Children from a pediatric renutrition center were prospectively enrolled in a fecal carriage study. Antibiotic exposure was recorded. ESBL-E strains were isolated using selective media from fecal samples obtained at admission and, when negative, also at discharge. ESBL-encoding genes were identified, their environments and plasmids were characterized, and clonality was assessed with polymerase chain reaction-based methods and pulsed-field gel electrophoresis for E. coli and Klebsiella pneumoniae. E. coli strains were subjected to multilocus sequence typing. RESULTS: The ESBL-E carriage rate was 31% at admission in the 55 children enrolled. All children enrolled received antibiotics during hospitalization. Among the ESBL-E-negative children, 16 were resampled at discharge, and the acquisition rate was 94%. The bla(CTX-M-15) gene was found in >90% of the carriers. Genetic environments and plasmid characterization evidenced the roles of a worldwide, previously described, multidrug-resistant region and of IncF plasmids in CTX-M-15 E. coli dissemination. Diversity of CTX-M-15-carrying genetic structures and clonality of acquired ESBL E. coli suggested horizontal genetic transfer and underlined the potential of some ST types for nosocomial cross-transmission. CONCLUSIONS: Cross-transmission and high selective pressure lead to very high acquisition of ESBL-E carriage, contributing to dissemination in the community. Strict hygiene measures as well as careful balancing of benefit-risk ratio of current antibiotic policies need to be reevaluated.

Study of canal sediments contaminated with heavy metals: fungal versus bacterial bioleaching techniques.

Filamentous fungi and lithotrophic bacteria were used to leach heavy metals from dredged sediments in semi-pilot scale air-lift bioreactors. A preliminary physico-chemical characterization of the sediments comprising a sequential extraction study revealed their high metallic contamination and a predominant association of the metals with sulphides and organic matter. The mobility of heavy metals from sediments was ranked by decreasing order as follows: Mn > Zn > Cd > Cu > Pb. The conditions that favoured the solubilization of heavy metals by filamentous fungi turned out to be also favourable for the activity of the sediment organotrophic bacteria. The latter produced organic acids under temporary hypoxic conditions and resulted in the solubilization of 77% of manganese, 44% of zinc, 12% of copper, and less than 2% of cadmium or lead. In general, the fungal organotrophic treatments were limited to the relatively mobile metals due to the weak nature of the organic acids produced and to their microbial consumption under limited saccharose conditions. The lithotrophic treatments yielded higher solubilization results than the organotrophic experiments. Sulphur resulted in a faster, and for some metals such as copper and cadmium, in better bioleaching results compared with reduced iron or with a combination of reduced iron and sulphur. The bioleaching percentages varied between 72 and 93% for cadmium, copper, manganese and zinc, except for lead because of the poor solubility of lead sulphate. The sediment's lithotrophic bacteria acidified the matrix through sulphur oxidation, and leached both loosely and tightly bound metals.

E-SovTox: An online database of the main publicly-available sources of toxicity data concerning REACH-relevant chemicals published in the Russian language.

A new open-access online database, E-SovTox, is presented. E-SovTox provides toxicological data for substances relevant to the EU Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) system, from publicly-available Russian language data sources. The database contains information selected mainly from scientific journals published during the Soviet Union era. The main information source for this database - the journal, Gigiena Truda i Professional'nye Zabolevania [Industrial Hygiene and Occupational Diseases], published between 1957 and 1992 - features acute, but also chronic, toxicity data for numerous industrial chemicals, e.g. for rats, mice, guinea-pigs and rabbits. The main goal of the abovementioned toxicity studies was to derive the maximum allowable concentration limits for industrial chemicals in the occupational health settings of the former Soviet Union. Thus, articles featured in the database include mostly data on LD50 values, skin and eye irritation, skin sensitisation and cumulative properties. Currently, the E-SovTox database contains toxicity data selected from more than 500 papers covering more than 600 chemicals. The user is provided with the main toxicity information, as well as abstracts of these papers in Russian and in English (given as provided in the original publication). The search engine allows cross-searching of the database by the name or CAS number of the compound, and the author of the paper. The E-SovTox database can be used as a decision-support tool by researchers and regulators for the hazard assessment of chemical substances.

Toxicity of nanoparticles of CuO, ZnO and TiO2 to microalgae Pseudokirchneriella subcapitata.

Toxicities of ZnO, TiO2 and CuO nanoparticles to Pseudokirchneriella subcapitata were determined using OECD 201 algal growth inhibition test taking in account potential shading of light. The results showed that the shading effect by nanoparticles was negligible. ZnO nanoparticles were most toxic followed by nano CuO and nano TiO2. The toxicities of bulk and nano ZnO particles were both similar to that of ZnSO4 (72 h EC50 approximately 0.04 mg Zn/l). Thus, in this low concentration range the toxicity was attributed solely to solubilized Zn2+ ions. Bulk TiO2 (EC50=35.9 mg Ti/l) and bulk CuO (EC50=11.55 mg Cu/l) were less toxic than their nano formulations (EC50=5.83 mg Ti/l and 0.71 mg Cu/l). NOEC (no-observed-effect-concentrations) that may be used for risk assessment purposes for bulk and nano ZnO did not differ (approximately 0.02 mg Zn/l). NOEC for nano CuO was 0.42 mg Cu/l and for bulk CuO 8.03 mg Cu/l. For nano TiO2 the NOEC was 0.98 mg Ti/l and for bulk TiO2 10.1 mg Ti/l. Nano TiO2 formed characteristic aggregates entrapping algal cells that may contribute to the toxic effect of nano TiO2 to algae. At 72 h EC50 values of nano CuO and CuO, 25% of copper from nano CuO was bioavailable and only 0.18% of copper from bulk CuO. Thus, according to recombinant bacterial and yeast Cu-sensors, copper from nano CuO was 141-fold more bioavailable than from bulk CuO. Also, toxic effects of Cu oxides to algae were due to bioavailable copper ions. To our knowledge, this is one of the first systematic studies on effects of metal oxide nanoparticles on algal growth and the first describing toxic effects of nano CuO towards algae.

Toxicity of nanosized and bulk ZnO, CuO and TiO2 to bacteria Vibrio fischeri and crustaceans Daphnia magna and Thamnocephalus platyurus.

As the production of nanoparticles of ZnO, TiO2 and CuO is increasing, their (eco)toxicity to bacteria Vibrio fischeri and crustaceans Daphnia magna and Thamnocephalus platyurus was studied with a special emphasis on product formulations (nano or bulk oxides) and solubilization of particles. Our innovative approach based on the combination of traditional ecotoxicology methods and metal-specific recombinant biosensors allowed to clearly differentiate the toxic effects of metal oxides per se and solubilized metal ions. Suspensions of nano and bulk TiO2 were not toxic even at 20 g l(-1). All Zn formulations were very toxic: L(E)C50 (mg l(-1)) for bulk ZnO, nanoZnO and ZnSO4.7H2O: 1.8, 1.9, 1.1 (V. fischeri); 8.8, 3.2, 6.1 (D. magna) and 0.24, 0.18, 0.98 (T. platyurus), respectively. The toxicity was due to solubilized Zn ions as proved with recombinant Zn-sensor bacteria. Differently from Zn compounds, Cu compounds had different toxicities: L(E)C50 (mg l(-1)) for bulk CuO, nano CuO and CuSO4: 3811, 79, 1.6 (V. fischeri), 165, 3.2, 0,17 (D. magna) and 95, 2.1, 0.11 (T. platyurus), respectively. Cu-sensor bacteria showed that toxicity to V. fischeri and T. platyurus was largely explained by soluble Cu ions. However, for Daphnia magna, nano and bulk CuO proved less bioavailable than for bacterial Cu-sensor. This is the first evaluation of ZnO, CuO and TiO2 toxicity to V. fischeri and T. platyurus. For nano ZnO and nano CuO this is also a first study for D. magna.

Biotests and Biosensors for Ecotoxicology of Metal Oxide Nanoparticles: A Minireview.

Nanotechnologies have become a significant priority worldwide. Several manufactured nanoparticles - particles with one dimension less than 100 nm - are increasingly used in consumer products. At nanosize range, the properties of materials differ substantially from bulk materials of the same composition, mostly due to the increased specific surface area and reactivity, which may lead to increased bioavailability and toxicity. Thus, for the assessment of sustainability of nanotechnologies, hazards of manufactured nanoparticles have to be studied. Despite all the above mentioned, the data on the potential environmental effects of nanoparticles are rare. This mini-review is summarizing the emerging information on different aspects of ecotoxicological hazard of metal oxide nanoparticles, focusing on TiO2, ZnO and CuO. Various biotests that have been successfully used for evaluation of ecotoxic properties of pollutants to invertebrates, algae and bacteria and now increasingly applied for evaluation of hazard of nanoparticles at different levels of the aquatic food-web are discussed. Knowing the benefits and potential drawbacks of these systems, a suite of tests for evaluation of environmental hazard of nanoparticles is proposed. Special attention is paid to the influence of particle solubility and to recombinant metal-sensing bacteria as powerful tools for quantification of metal bioavailability. Using recombinant metal-specific bacterial biosensors and multitrophic ecotoxicity assays in tandem will create new scientific knowledge on the respective role of ionic species and of particles in toxicity of metal oxide nanoparticles.

Rapid screening for soil ecotoxicity with a battery of luminescent bacteria tests.

A bacterial test battery, involving i) Microtox, an aquatic test, ii) the Flash assay, a soil-suspension test (with Vibrio fischeri as the test organism), and iii) the Metal Detector assay, a semi-specific aquatic test for heavy metals (with recombinant luminescent Escherichia coli), was used in a combined toxicological and chemical hazard assessment of Estonian soils sampled from a former Soviet military airfield (13 samples) and from traffic-influenced roadsides (5 samples). The soils showed slightly elevated levels of total petroleum hydrocarbons (TPH), but not of heavy metals. In most of the samples, the levels of TPH did not exceed the Estonian permitted limit values set for residential areas. Toxicity testing was performed on both fresh and dried soils, after aqueous extraction for 1 hour and 24 hours. The toxicity results obtained with the Microtox test did not significantly differ in all of the sample treatment schemes; however, it appeared that the drying and sieving of the soils increased the bioavailability of toxicants, probably due to an enlarged reactive soil surface area. According to chemical analysis of the soils and the data from the Microtox test and the Metal Detector assay (performed on aqueous elutriates of the soils), these soils would not be considered to be hazardous. In contrast, the Flash assay performed on soil-water suspensions of dried soils, showed that most of the soils were toxic and thus probably contained undetermined particle-bound bioavailable toxicants. The photobacterial toxicity test (the Flash assay) can be recommended for the rapid screening of soils, as it is sensitive, cheap and inexpensive, and provides valuable information on particle-bound bioavailable toxicants, useful for complementing a chemical analysis and for assessing the risks originating from polluted soils.

Biotests and biosensors in ecotoxicological risk assessment of field soils polluted with zinc, lead, and cadmium.

The combined chemical and ecotoxicological hazard evaluation study was conducted on 60 smelter-influenced soils containing 1 to 13, 50 to 653, and 100 to 1,198 mg/kg of Cd, Pb, and Zn, respectively. For these soils (liquid-to-soil ratio = 10), water extractability of Zn, Cd, and Pb was less than 0.19% (median values). Acetic acid (0.11 M) extracted 23, 9.7, and 0.7% of Cd, Zn, and Pb, respectively. Although heavy metal concentrations in the studied soils were high, the toxic effects of water extracts were observed only in few samples and in few biotests (algae Selenastrum capricornutum and metal detector assay). For most of the aquatic test organisms (e.g., crustaceans, photobacteria), the bioavailable concentrations of metals in soil-water extracts were either subtoxic, or the adverse effects were compensated by soil nutrients, etc. However, analysis of the soils with recombinant Cd sensor Bacillus subtilis (pTOO24) showed that about 65% of these apparently subtoxic samples contained bioavailable Cd when analyzed in the suspension assay (detection limit 1.5 mg Cd/kg soil), indicating the desorption of Cd induced by direct contact of bacteria with soil particles. The median bioavailable fraction of Cd (1%) was 23-fold lower than the fraction extracted by acetic acid. The Pb-Cd sensor Staphylococcus aureus (pT0024) detected bioavailable Pb only in the suspensions of five of the most lead-polluted soils (>417 mg Pb/kg): the median bioavailability of Pb was 0.42%. Consequently, the hazard assessment relying on total metal levels in soils should be revised by critical comparison with data obtained from bioassays. Development and use of biosensors (excellent tools for mechanistic studies and signaling hazard already at subtoxic level) should be encouraged.

Recombinant luminescent bacterial sensors for the measurement of bioavailability of cadmium and lead in soils polluted by metal smelters.

Environmental hazard of heavy metals in soils depends to a large extent on their bioavailability. The approach used in this study enables the determination of bioavailable metals in solid-phase samples. Two recombinant bacterial sensors, one responding specifically to cadmium and the other to lead and cadmium by increase of luminescence (firefly luciferase was used as a reporter) were used to determine the bioavailability of these metals in soil-water suspensions (a contact assay) and respective particle-free extracts. Fifty agricultural soils sampled near zinc and lead smelters in the Northern France containing up to (mg/kg) 20.1 of Cd, 1050 of Pb and 1390 of Zn were analysed. As the soil matrix interferes with the assay, recombinant luminescent control bacteria lacking the metal recognizing protein and corresponding promoter (thus, being not metal-inducible) but otherwise comparable to the sensor bacteria (the same host bacterium and plasmid encoding luciferase) were used in parallel to take into account the possible quenching and/or stimulating effects of the sample on the luminescence of the sensor bacteria. Both, chemical and sensor analysis showed that only microg/l levels of metals were extracted from the soil into the water phase (0.1% of the total Cd, 0.07% of Pb and 0.5% of Zn). However, 115-fold more Cd and 40-fold more Pb proved bioavailable if the sensor bacteria were incubated in soil suspensions (i.e., in the contact assay). The bioavailability of metals in different soils varied (depending probably on soil type) ranging from 0.5% to 56% for cadmium and from 0.2% to 8.6% for lead.

Toxicity of 58 substituted anilines and phenols to algae Pseudokirchneriella subcapitata and bacteria Vibrio fischeri: comparison with published data and QSARs.

A congeneric set of 58 substituted anilines and phenols was tested using the 72-h algal growth inhibition assay with Pseudokirchneriella subcapitata and 15-min Vibrio fischeri luminescence inhibition assay. The set contained molecules substituted with one, two or three groups chosen from -chloro, -methyl or -ethyl. For 48 compounds there was no REACH-compatible algal toxicity data available before. The experimentally obtained EC50 values (mg L(-1)) for algae ranged from 1.43 (3,4,5-trichloroaniline) to 197 (phenol) and for V. fischeri from 0.37 (2,3,5-trichlorophenol) to 491 (aniline). Only five of the tested 58 chemicals showed inhibitory effect to algae at concentrations >100 mg L(-1), i.e. could be classified as "not harmful", 32 chemicals as "harmful" (10-100 mg L(-1)) and 21 as "toxic" (1-10 mg L(-1)). The occupied para-position tended to increase toxicity whereas most of the ortho-substituted congeners were the least toxic. As a rule, the higher the number of substituents the higher the hydrophobicity and toxicity. However, in case of both assays, the compounds of similar hydrophobicity showed up to 30-fold different toxicities. There were also assay/organism dependent tendencies: phenols were more toxic than anilines in the V. fischeri assay but not in the algal test. The comparison of the experimental toxicity data to the data available from the literature as well as to QSAR predictions showed that toxicity of phenols to algae can be modeled based on hydrophobicity, whereas the toxicity of anilines to algae as well as toxicity of both anilines and phenols to V. fischeri depended on other characteristics in addition to log K(ow).

Particle-size effect of CuO and ZnO on biogas and methane production during anaerobic digestion.

The effects of bulk- and nano-sized CuO and ZnO particles on biogas and methane production during anaerobic digestion of cattle manure were studied for a period of 14 days at 36 °C using the ISO 13641-2 guidelines. Biogas production was severely affected at concentrations of bulk and nanoparticles over 120 and 15 mg/L for CuO and 240 and 120 mg/L for ZnO, respectively. EC50 concentrations for methane inhibition were estimated to be 129 mg Cu/L for bulk CuO, 10.7 mg Cu/L for nano CuO, 101 mg Zn/L for bulk ZnO and 57.4 mg Zn/L for nano ZnO. The solubility of CuO nanoparticles in the reaction mixture was observed after 14 days of incubation and was significantly higher than the levels observed for ZnO. These results are of significant importance, as it is the first time that the effects of metal oxide particle size on biogas and methane production have been studied.

Changes in the Daphnia magna midgut upon ingestion of copper oxide nanoparticles: a transmission electron microscopy study.

This work is a follow-up of our previous paper (Heinlaan et al., 2008. Chemosphere 71, 1308-1316) where we showed about 50-fold higher acute toxicity of CuO nanoparticles (NPs) compared to bulk CuO to water flea Daphnia magna. In the current work transmission electron microscopy (TEM) was used to determine potential time-dependent changes in D. magna midgut epithelium ultrastructure upon exposure to CuO NPs compared to bulk CuO at their 48 h EC(50) levels: 4.0 and 175 mg CuO/L, respectively. Special attention was on potential internalization of CuO NPs by midgut epithelial cells. Ingestion of both CuO formulations by daphnids was evident already after 10 min of exposure. In the midgut lumen CuO NPs were dispersed whereas bulk CuO was clumped. By the 48th hour of exposure to CuO NPs (but not to equitoxic concentrations of bulk CuO) the following ultrastructural changes in midgut epithelium of daphnids were observed: protrusion of epithelial cells into the midgut lumen, presence of CuO NPs in circular structures analogous to membrane vesicles from holocrine secretion in the midgut lumen. Implicit internalization of CuO NPs via D. magna midgut epithelial cells was not evident however CuO NPs were no longer contained within the peritrophic membrane but located between the midgut epithelium microvilli. Interestingly, upon exposure to CuO NPs bacterial colonization of the midgut occurred. Ultrastructural changes in the midgut of D. magna upon exposure to CuO NPs but not to bulk CuO refer to its nanosize-related adverse effects. Time-dependent solubilisation of CuO NPs and bulk CuO in the test medium was quantified by recombinant Cu-sensor bacteria: by the 48th hour of exposure to bulk CuO, the concentration of solubilised copper ions was 0.05 ± 0.01 mg Cu/L that was comparable to the acute EC(50) value of Cu-ions to D. magna (48 h CuSO(4) EC(50) = 0.07 ± 0.01 mg Cu/L). However, in case of CuO NPs, the solubilised Cu-ions 0.01 ± 0.001 mg Cu/L, explained only part of the toxicity.

From ecotoxicology to nanoecotoxicology.

For hazard assessment of NPs quantitative nanoecotoxicological data are required. The objective of this review was to evaluate the currently existing literature data on toxicity (L(E)C50 values) of synthetic NPs in environmentally relevant species in order to: (i) identify tentatively most harmful NPs and most sensitive organism groups, and (ii) to provide relevant ecotoxicological information for further risk assessment. The focus was set on selected synthetic NPs (nano TiO(2), nano ZnO, nano CuO, nano Ag, SWCNTs, MWCNs and C60-fullerenes) and organism groups representing main food-chain levels (bacteria, algae, crustaceans, ciliates, fish, yeasts and nematodes). Altogether 77 effect values were found, mostly for nano TiO(2) (31%), C60 (18%), nano ZnO (17%), nano Ag (13%), SWCNTs and nano CuO (both 9%). Only 3% of the available quantitative ecotoxicological information concerned MWCNTs. Organism-wise, 33% of the data concerned crustaceans, 27% bacteria, 14% algae and 13% fish. For all organism groups studied, solubility of CuO- and ZnO-NPs was a key factor in their aquatic toxicity. On the basis of the 34 median L(E)C50 values derived from 77 individual values, NPs were ranked according to their lowest median L(E)C50 value for the above described organism groups: the most harmful were nano Ag and nano ZnO that were classified "extremely toxic", (L(E)C50<0.1mg/l), followed by C60 fullerenes and nano CuO that were classified "very toxic", (L(E)C50 0.1-1mg/l). SWCNTs and MWCNTs were classified "toxic" (L(E)C50 1-10mg/l). Nano TiO(2) was classified as "harmful", (L(E)C50 10-100mg/l). Throughout, algae and crustaceans (daphnids) were most sensitive and thus probably most vulnerable organism groups in aquatic exposure to NPs. Very low L(E)C50 values should deserve thorough attention of environmental risk assessors for evaluation of the potential adverse effects of synthetic NPs on ecosystems. As the quantitative nanoecotoxicological data are still rare, further studies are needed.

Toxicological information on chemicals published in the Russian language: Contribution to REACH and 3Rs.

This review is reporting on the current situation of publicly available toxicological and ecotoxicological information on chemicals published in Russian language in various libraries, databases as well as in the Internet. This information can be beneficial for the new EU chemical policy REACH and for the development of intelligent testing strategies (involving also QSAR and QAAR) that enable a significant increase in the use of non-testing information for regulatory decision making, thus minimizing the need for animal testing according to the 3R's strategy. Currently, the access to this information is limited due to the language barrier and low level of digitalization of respective journals and books. Fortunately, on-line translation services are overcoming language barriers already now.

Toxicity of nanoparticles of ZnO, CuO and TiO2 to yeast Saccharomyces cerevisiae.

The aim of this study was to evaluate the toxic effect of nanosized ZnO, CuO and TiO(2) to Saccharomyces cerevisiae - a widely used unicellular eukaryotic model organisms in molecular and cell biology. The effect of metal oxide nanoparticles, their bulk forms and respective ionic forms were compared. The bioavailable Zn(2+) and Cu(2+) ions in the growth medium were quantified by recombinant microbial sensors. Nano and bulk TiO(2) were not toxic even at 20000 mg/l. Both, nano and bulk ZnO were of comparable toxicity (8-h EC(50) 121-134 mg ZnO/l and 24-h EC(50) 131-158 mg/l). The toxicity was explained by soluble Zn-ions as proved by the microbial sensor. However, nano CuO was about 60-fold more toxic than bulk CuO: 8-h EC(50) were 20.7 and 1297 mg CuO/l and 24-h EC(50) were 13.4 and 873 mg/l, respectively. The increase in toxicity of both CuO formulations at 24th hour of growth was due to the increased dissolution of copper ions from CuO over time. Comparison of EC(50) values of nano CuO, bulk CuO and Cu(2+) with bioavailable copper concentrations in the growth medium showed that the solubilized Cu-ions explained only about 50% of the toxicity of both, nano and bulk CuO. To our knowledge, this is the first study that evaluates the toxicity of ZnO, CuO and TiO(2) nanoparticles to S.cerevisiae.

Toxicity testing of heavy-metal-polluted soils with algae Selenastrum capricornutum: a soil suspension assay.

A small-scale Selenastrum capricornutum (Rhapidocelis subcapitata) growth inhibition assay was applied to the toxicity testing of suspensions of heavy-metal-polluted soils. The OECD 201 standard test procedure was followed, and algal biomass was measured by the fluorescence of extracted chlorophyll. The soils, which contained up to (per kilogram) 1390 mg of Zn, 20 mg of Cd, and 1050 mg of Pb were sampled around lead and zinc smelters in northern France. The water extractability of the metals in suspensions (1 part soil/99 parts water w/v) was not proportional to the pollution level, as extractability was lower for soil samples that were more polluted. Thus, the same amount of metals could be leached out of soils of different levels of pollution, showing that total concentrations of heavy metals in soil (currently used for risk assessment purposes) are poor predictors of the real environmental risk via the soil-water path. Despite high concentrations of water-extracted zinc (0.6-1.4 mg/L of Zn in the test), exceeding by approximately 10-fold the EC(50) value for S. capricornutum (0.1 mg Zn/L), 72-h algal growth in the soil extracts was comparable or better than growth in the standard control OECD mineral medium. The soil suspension stimulated the growth of algae up to eightfold greater than growth using the OECD control medium. Growth stimulation of algae was observed even when soil suspensions contained up to 12.5 mg Zn/L and could not be explained by supplementary nitrogen, phosphorous, and carbonate leached from the soil. However, if the growth of algae in suspensions of clean and polluted soils was compared, a dose-dependent inhibitory effect of metals on algal growth was demonstrated. Thus, as soil contains nutrients/supplements that mask the adverse effect of heavy metals, a clean soil that has properties similar to the polluted soils should be used instead of mineral salt solution as a control for analysis of the ecotoxicity of soils.