Aminocatalysts are More Environmentally Friendly than Hydrogen-Bonding Catalysts.

The importance of asymmetric organocatalysis in contemporary organic synthesis is widely acknowledged. However, there are practically no data on the environmental safety of organocatalysts, although this aspect is crucial for the sustainability of all new materials, chemicals, and technologies. To start to fill this data-gap, a library of 26 organocatalysts containing hydrogen-bonding catalysts [(thio)ureas and squaramides] and aminocatalysts (primary or secondary amines) was evaluated for their toxicity using the naturally luminescent Vibrio fischeri bacteria (ISO assay; one of the most widely used ecotoxicity tests). Thioureas and squaramides were shown to be relatively toxic: none of them was ranked as "not harmful" (i. e., half maximal effective concentration EC50 >100 mg L-1 ), whereas the presence of the trifluoromethyl moiety increased their toxic effect. Importantly, the aminocatalysts, whose EC50 values ranged from 25 to >300 mg L-1 , could be considered remarkably more environmentally safe or green alternatives.

Antibacterial Activity of Positively and Negatively Charged Hematite (α-Fe2O3) Nanoparticles to Escherichia coli, Staphylococcus aureus and Vibrio fischeri.

In the current study, the antibacterial activity of positively and negatively charged spherical hematite (α-Fe2O3) nanoparticles (NPs) with primary size of 45 and 70 nm was evaluated against clinically relevant bacteria Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive) as well as against naturally bioluminescent bacteria Vibrio fischeri (an ecotoxicological model organism). α-Fe2O3 NPs were synthesized using a simple green hydrothermal method and the surface charge was altered via citrate coating. To minimize the interference of testing environment with NP's physic-chemical properties, E. coli and S. aureus were exposed to NPs in deionized water for 30 min and 24 h, covering concentrations from 1 to 1000 mg/L. The growth inhibition was evaluated following the postexposure colony-forming ability of bacteria on toxicant-free agar plates. The positively charged α-Fe2O3 at concentrations from 100 mg/L upwards showed inhibitory activity towards E. coli already after 30 min of contact. Extending the exposure to 24 h caused total inhibition of growth at 100 mg/L. Bactericidal activity of positively charged hematite NPs against S. aureus was not observed up to 1000 mg/L. Differently from positively charged hematite NPs, negatively charged citrate-coated α-Fe2O3 NPs did not exhibit any antibacterial activity against E. coli and S. aureus even at 1000 mg/L. Confocal laser scanning microscopy and flow cytometer analysis showed that bacteria were more tightly associated with positively charged α-Fe2O3 NPs than with negatively charged citrate-coated α-Fe2O3 NPs. Moreover, the observed associations were more evident in the case of E. coli than S. aureus, being coherent with the toxicity results. Vibrio fischeri bioluminescence inhibition assays (exposure medium 2% NaCl) and colony forming ability on agar plates showed no (eco)toxicity of α-Fe2O3 (EC50 and MBC > 1000 mg/L).

Hazard evaluation of polystyrene nanoplastic with nine bioassays did not show particle-specific acute toxicity.

Plastic is a wide-spread pollutant and must be evaluated for potential adverse effects of its breakdown product, microplastic (≤5 mm) along with its subfraction, nanoplastic (1-100 nm). Risk assessment of pollutants cannot be conducted without their toxicity (dose-response) data. In this study, toxicity of polystyrene nanoplastics (PS-NPL) was evaluated using 8 acute and 1 subchronic toxicity assays with 10 organisms of different biological complexity (bacteria, yeast, algae, protozoans, mammalian cells in vitro, crustaceans, midge larvae). Commercial 26 and 100 nm carboxylated PS-NPL spheres were chosen as model and tested in nominal concentrations up to 100 mg/L (1.025·1016 26 nm and 1.83·1014 100 nm particles/L). In most of the assays, both PS-NPL proved non-toxic (L(E)C50 > 100 mg/L) but three tests (V. fischeri, R. subcapitata, D. magna) flagged toxicity in 'as received' 26 nm PS-NPL and D. magna also in 100 nm PS-NPL (EC50 ranging from 13 to 71 mg/L). As, according to manufacturers, both PS-NPL suspensions contained additives (surfactants and biocidal NaN3), the three toxicity tests were repeated also on dialysed PS-NPL and on NaN3. Non-toxicity of dialysed PS-NPL indicated that the toxicity of 'as-received' PS-NPL was not particle-specific but false positive due to water-soluble additives in the PS-NPL preparations. NaN3 was very toxic to D. magna (48 h EC50 = 0.05 ± 0.03 mg NaN3/L), toxic to R. subcapitata (72 h EC50 = 4.97 ± 3.7 mg NaN3/L) and non-toxic to V. fischeri. Toxicity of 'as-received' PS-NPL was not fully explainable by NaN3 but also attributable to other additives in the suspensions. Toxicity research of microplastic using commercial model particles must always consider the potential influence of additives, e.g. test the toxicity of dialysed NPL for comparison. In our study, D. magna, R. subcapitata and V. fischeri were the most sensitive to PS-NPL water-soluble additives and flagged their presence in NPL preparations.

Toxicity profiling of 24 l-phenylalanine derived ionic liquids based on pyridinium, imidazolium and cholinium cations and varying alkyl chains using rapid screening Vibrio fischeri bioassay.

A library of 24 pyridinium-, imidazolium-, and cholinium-based ionic liquids (ILs) with varying alkyl chain from C2 to C16 was toxicologically profiled using naturally luminescent marine bacteria Vibrio fischeri. The toxicity (30-min EC50) of studied ILs to Vibrio fischeri ranged from 7.82 µM (4.2 mg/L) (PyC12Phe) to 3096 µM (1227 mg/L) (ImidC2Phe), i.e. from "toxic" (EC50 1-10 mg/L) to "not harmful" (EC50 > 100 mg/L). Inhibition of the bacterial luminescence upon 30-min exposure to ILs correlated well with bacterial viability (exposure for 4 h). The toxicity of studied ILs was largely driven by the length of the alkyl chain (hydrophobicity) and not the type of cationic part of the IL: starting from C10 all the ILs irrespective of the cationic part proved "toxic". The toxicity of the studied ILs was increasing in parallel to their hydrophobicity up to log Kow = 1 (C8-C10) and then levelling up, being consistent with the previously obtained analogous data sets. The "cut-off" effect reported in this study for longer chain length members of the ILs series leads to the "limit" toxicity level for this type of ILs to be ca. 8 mM. Two open-access online tools (www.molinspiration.com and www.vcclab.org) have been applied for the calculation of the Kow values for the 24 ILs reported in this study and 21 ILs reported in the literature. This lead to plotting two nonlinear monotonic correlations between the values of experimental log (1/EC50) and calculated log Kow. The limitation of the online tools and an effect of the ILs structure on the "cut-off" effect have been discussed. The challenge of developing low microbial toxicity surface active ILs remains a significant task to overcome. Our results shed light on the new approaches for designing environmentally benign ILs and functional surfactants. As the hydrophobicity of the ILs significantly correlated with the toxicity, the Vibrio fischeri assay could be considered a powerful tool in providing toxicity data for building and evaluating the QSAR toxicity models for ILs.

Plasma membrane is the target of rapid antibacterial action of silver nanoparticles in Escherichia coli and Pseudomonas aeruginosa.

INTRODUCTION: Silver nanoparticles (AgNP) are widely used in consumer products and in medicine, mostly due to their excellent antimicrobial properties. One of the generally accepted antibacterial mechanisms of AgNP is their efficient contact with cells and dissolution in the close vicinity of bacterial cell envelope. Yet, the primary mechanism of cell wall damage and the events essential for bactericidal action of AgNP are not elucidated. MATERIALS AND METHODS: In this study we used a combination of various assays to differentiate the adverse effects of AgNP on bacterial cell envelope: outer membrane (OM) and plasma membrane (PM). RESULTS: We showed that PM was the main target of AgNP in gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa: AgNP depolarized PM, induced the leakage of the intracellular K+, and inhibited cellular respiration. The results of bacterial bioluminescence inhibition assay in combination with AgNP dissolution and oxidation assays demonstrated that the adverse effects of AgNP occurred at concentrations 7-160 µM. These toxic effects occurred already within the first few seconds of contact of bacteria and AgNP and were driven by dissolved Ag+ ions targeting bacterial PM. However, the irreversible inhibition of bacterial growth detected after 1-hour exposure occurred at 40 µM AgNP for P. aeruginosa and at 320 µM AgNP for E. coli. In contrast to effects on PM, AgNP and Ag+ ions had no significant effect on the permeability and integrity of bacterial OM, implying that AgNP indeed targeted mainly PM via dissolved Ag+ ions. CONCLUSION: AgNP exhibited antibacterial properties via rapid release of Ag+ ions targeting the PM and not the OM of gram-negative bacteria.

Toxicity of Nine (Doped) Rare Earth Metal Oxides and Respective Individual Metals to Aquatic Microorganisms Vibrio fischeri and Tetrahymena thermophila.

Despite the increasing use of rare earth elements (REEs) and oxides (REOs) in various technologies, the information on their ecotoxicological hazard is scarce. Here, the effects of La3+, Ce3+, Pr3+, Nd3+, Gd3+, CeO2, and eight doped REOs to marine bacteria Vibrio fischeri and freshwater protozoa Tetrahymena thermophila were studied in parallel with REO dopant metals (Co2+, Fe3+, Mn2+, Ni2+, Sr2+). The highest concentrations of REOs tested were 100 mg/L with protozoa in deionized water and 500 mg/L with bacteria in 2% NaCl. Although (i) most REOs produced reactive oxygen species; (ii) all studied soluble REEs were toxic to bacteria (half-effective concentration, EC50 3.5-21 mg metal/L; minimal bactericidal concentration, MBC 6.3-63 mg/L) and to protozoa (EC50 28-42 mg/L); and (iii) also some dopant metals (Ni2+, Fe3+) proved toxic (EC50 ≤ 3 mg/L), no toxicity of REOs to protozoa (EC50 > 100 mg/L) and bacteria (EC50 > 500 mg/L; MBC > 500 mg/L) was observed except for La2NiO4 (MBC 25 mg/L). According to kinetics of V. fischeri bioluminescence, the toxicity of REEs was triggered by disturbing cellular membrane integrity. Fortunately, as REEs and REOs are currently produced in moderate amounts and form in the environment insoluble salts and/or oxides, they apparently present no harm to aquatic bacteria and protozoa.

An Integrated Data-Driven Strategy for Safe-by-Design Nanoparticles: The FP7 MODERN Project.

The development and implementation of safe-by-design strategies is key for the safe development of future generations of nanotechnology enabled products. The safety testing of the huge variety of nanomaterials that can be synthetized is unfeasible due to time and cost constraints. Computational modeling facilitates the implementation of alternative testing strategies in a time and cost effective way. The development of predictive nanotoxicology models requires the use of high quality experimental data on the structure, physicochemical properties and bioactivity of nanomaterials. The FP7 Project MODERN has developed and evaluated the main components of a computational framework for the evaluation of the environmental and health impacts of nanoparticles. This chapter describes each of the elements of the framework including aspects related to data generation, management and integration; development of nanodescriptors; establishment of nanostructure-activity relationships; identification of nanoparticle categories; hazard ranking and risk assessment.

Multilaboratory evaluation of 15 bioassays for (eco)toxicity screening and hazard ranking of engineered nanomaterials: FP7 project NANOVALID.

Within EU FP7 project NANOVALID, the (eco)toxicity of 7 well-characterized engineered nanomaterials (NMs) was evaluated by 15 bioassays in 4 laboratories. The highest tested nominal concentration of NMs was 100 mg/l. The panel of the bioassays yielded the following toxicity order: Ag > ZnO > CuO > TiO2 > MWCNTs > SiO2 > Au. Ag, ZnO and CuO proved very toxic in the majority of assays, assumingly due to dissolution. The latter was supported by the parallel analysis of the toxicity of respective soluble metal salts. The most sensitive tests/species were Daphnia magna (towards Ag NMs, 24-h EC50 = 0.003 mg Ag/l), algae Raphidocelis subcapitata (ZnO and CuO, 72-h EC50 = 0.14 mg Zn/l and 0.7 mg Cu/l, respectively) and murine fibroblasts BALB/3T3 (CuO, 48-h EC50 = 0.7 mg Cu/l). MWCNTs showed toxicity only towards rat alveolar macrophages (EC50 = 15.3 mg/l) assumingly due to high aspect ratio and TiO2 towards R. subcapitata (EC50 = 6.8 mg Ti/l) due to agglomeration of TiO2 and entrapment of algal cells. Finally, we constructed a decision tree to select the bioassays for hazard ranking of NMs. For NM testing, we recommend a multitrophic suite of 4 in vitro (eco)toxicity assays: 48-h D. magna immobilization (OECD202), 72-h R. subcapitata growth inhibition (OECD201), 30-min Vibrio fischeri bioluminescence inhibition (ISO2010) and 48-h murine fibroblast BALB/3T3 neutral red uptake in vitro (OECD129) representing crustaceans, algae, bacteria and mammalian cells, respectively. Notably, our results showed that these assays, standardized for toxicity evaluation of "regular" chemicals, proved efficient also for shortlisting of hazardous NMs. Additional assays are recommended for immunotoxicity evaluation of high aspect ratio NMs (such as MWCNTs).

Toxicity of Water Accommodated Fractions of Estonian Shale Fuel Oils to Aquatic Organisms.

Estonia is the worldwide leading producer of the fuel oils from the oil shale. We evaluated the ecotoxicity of water accommodated fraction (WAF) of two Estonian shale fuel oils ("VKG D" and "VKG sweet") to aquatic species belonging to different trophic levels (marine bacteria, freshwater crustaceans and aquatic plants). Artificial fresh water and natural lake water were used to prepare WAFs. "VKG sweet" (lower density) proved more toxic to aquatic species than "VKG D" (higher density). Our data indicate that though shale oils were very toxic to crustaceans, the short-term exposure of Daphnia magna to sub-lethal concentrations of shale fuel oils WAFs may increase the reproductive potential of survived organisms. The weak correlation between measured chemical parameters (C10-C40 hydrocarbons and sum of 16 PAHs) and WAF's toxicity to studied species indicates that such integrated chemical parameters are not very informative for prediction of shale fuel oils ecotoxicity.

An interlaboratory comparison of nanosilver characterisation and hazard identification: Harmonising techniques for high quality data.

Within the FP7 EU project NanoValid a consortium of six partners jointly investigated the hazard of silver nanoparticles (AgNPs) paying special attention to methodical aspects that are important for providing high-quality ecotoxicity data. Laboratories were supplied with the same original stock dispersion of AgNPs. All partners applied a harmonised procedure for storage and preparation of toxicity test suspensions. Altogether ten different toxicity assays with a range of environmentally relevant test species from different trophic levels were conducted in parallel to AgNP characterisation in the respective test media. The paper presents a comprehensive dataset of toxicity values and AgNP characteristics like hydrodynamic sizes of AgNP agglomerates and the share (%) of Ag(+)-species (the concentration of Ag(+)-species in relation to the total measured concentration of Ag). The studied AgNP preparation (20.4±6.8 nm primary size, mean total Ag concentration 41.14 mg/L, 46-68% of soluble Ag(+)-species in stock, 123.8±12.2 nm mean z-average value in dH2O) showed extreme toxicity to crustaceans Daphnia magna, algae Pseudokirchneriella subcapitata and zebrafish Danio rerio embryos (EC50<0.01 mg total Ag/L), was very toxic in the in vitro assay with rainbow trout Oncorhynchus mykiss gut cells (EC50: 0.01-1 mg total Ag/L); toxic to bacteria Vibrio fischeri, protozoa Tetrahymena thermophila (EC50: 1-10 mg total Ag/L) and harmful to marine crustaceans Artemia franciscana (EC50: 10-100 mg total Ag/L). Along with AgNPs, also the toxicity of AgNO3 was analyzed. The toxicity data revealed the same hazard ranking for AgNPs and AgNO3 (i.e. the EC50 values were in the same order of magnitude) proving the importance of soluble Ag(+)-species analysis for predicting the hazard of AgNPs. The study clearly points to the need for harmonised procedures for the characterisation of NMs. Harmonised procedures should consider: (i) measuring the AgNP properties like hydrodynamic size and metal ions species in each toxicity test medium at a range of concentrations, and (ii) including soluble metal salt control both in toxicity testing as well as in Ag(+)-species measurements. The present study is among the first nanomaterial interlaboratory comparison studies with the aim to improve the hazard identification testing protocols.

Toxicity of 11 Metal Oxide Nanoparticles to Three Mammalian Cell Types In Vitro.

The knowledge on potential harmful effects of metallic nanomaterials lags behind their increased use in consumer products and therefore, the safety data on various nanomaterials applicable for risk assessment are urgently needed. In this study, 11 metal oxide nanoparticles (MeOx NPs) prepared using flame pyrolysis method were analyzed for their toxicity against human alveolar epithelial cells A549, human epithelial colorectal cells Caco2 and murine fibroblast cell line Balb/c 3T3. The cell lines were exposed for 24 h to suspensions of 3-100 µg/mL MeOx NPs and cellular viability was evaluated using. Neutral Red Uptake (NRU) assay. In parallel to NPs, toxicity of soluble salts of respective metals was analyzed, to reveal the possible cellular effects of metal ions shedding from the NPs. The potency of MeOx to produce reactive oxygen species was evaluated in the cell-free assay. The used three cell lines showed comparable toxicity responses to NPs and their metal ion counterparts in the current test setting. Six MeOx NPs (Al2O3, Fe3O4, MgO, SiO2, TiO2, WO3) did not show toxic effects below 100 µg/mL. For five MeOx NPs, the averaged 24 h IC50 values for the three mammalian cell lines were 16.4 µg/mL for CuO, 22.4 µg/mL for ZnO, 57.3 µg/mL for Sb2O3, 132.3 µg/mL for Mn3O4 and 129 µg/mL for Co3O4. Comparison of the dissolution level of MeOx and the toxicity of soluble salts allowed to conclude that the toxicity of CuO, ZnO and Sb2O3 NPs was driven by release of metal ions. The toxic effects of Mn3O4 and Co3O4 could be attributed to the ROS-inducing ability of these NPs. All the NPs were internalized by the cells according to light microscopy studies but also proven by TEM, and internalization of Co3O4 NPs seemed to be most prominent in this aspect. In conclusion, this work provides valuable toxicological data for a library of 11 MeOx NPs. Combining the knowledge on toxic or non-toxic nature of nanomaterials may be used for safe-by-design approach.

A novel method for comparison of biocidal properties of nanomaterials to bacteria, yeasts and algae.

Toxicity testing of nanomaterials (NMs) is experimentally challenging because NMs may interfere with test environment and assay components. In this work we propose a simple and reliable method--a 'spot test' to compare biocidal potency of NMs to unicellular microorganisms such as bacteria, yeasts and algae. The assay is straightforward: cells are incubated in deionized water suspensions of NMs for up to 24h and then pipetted as a 'spot' on agarized medium. Altogether seven bacterial strains, yeast and a microalga were tested. CuO, TiO2 and two different Ag NPs, multi-wall C-nanotubes (MWCNTs), AgNO3, CuSO4, 3,5-dichlorophenol, triclosan and H2O2 were analyzed. The biocidal potency of tested substances ranged from 0.1mg/L to >1000 mg/L; whereas, the least potent NMs toward all test species were TiO2 NPs and MWCNTs and most potent Ag and CuO NPs. Based on the similar toxicity pattern of the tested chemicals on the nine unicellular organisms in deionized water we conclude that toxicity mechanism of biocidal chemicals seems to be similar, whatever the organism (bacteria, yeast, alga). Therefore, when the organisms are not 'protected' by their environment that usually includes various organic and inorganic supplements their tolerance to toxicants is chemical- rather than organism-dependent.

Environmental effects of soil contamination by shale fuel oils.

Estonia is currently one of the leading producers of shale oils in the world. Increased production, transportation and use of shale oils entail risks of environmental contamination. This paper studies the behaviour of two shale fuel oils (SFOs)--'VKG D' and 'VKG sweet'--in different soil matrices under natural climatic conditions. Dynamics of SFOs' hydrocarbons (C10-C40), 16 PAHs, and a number of soil heterotrophic bacteria in oil-spiked soils was investigated during the long-term (1 year) outdoor experiment. In parallel, toxicity of aqueous leachates of oil-spiked soils to aquatic organisms (crustaceans Daphnia magna and Thamnocephalus platyurus and marine bacteria Vibrio fischeri) and terrestrial plants (Sinapis alba and Hordeum vulgare) was evaluated. Our data showed that in temperate climate conditions, the degradation of SFOs in the oil-contaminated soils was very slow: after 1 year of treatment, the decrease of total hydrocarbons' content in the soil did not exceed 25 %. In spite of the comparable chemical composition of the two studied SFOs, the VKG sweet posed higher hazard to the environment than the heavier fraction (VKG D) due to its higher mobility in the soil as well as higher toxicity to aquatic and terrestrial species. Our study demonstrated that the correlation between chemical parameters (such as total hydrocarbons or total PAHs) widely used for the evaluation of the soil pollution levels and corresponding toxicity to aquatic and terrestrial organisms was weak.

Measurement of baseline toxicity and QSAR analysis of 50 non-polar and 58 polar narcotic chemicals for the alga Pseudokirchneriella subcapitata.

In this paper a set of homogenous experimental algal toxicity data was measured for 50 non-polar narcotic chemicals using the alga Pseudokirchneriella subcapitata in a closed test with a growth rate endpoint. Most of the tested compounds are high volume industrial chemicals that so far lacked published REACH-compliant algal growth inhibition values. The test protocol fulfilled the criteria set forth in the OECD guideline 201 and had the same sensitivity as the open test which allowed direct comparison of toxicity values. Baseline QSAR model for non-polar narcotic compounds was established and compared with previous analogous models. Multi-linear QSAR model was derived for the non-polar and 58 previously tested polar (anilines and phenols) narcotic compounds modulating hydrophobicity, molecular size, electronic and molecular stability effects coded in the molecular descriptors. Descriptors in the model were analyzed and applicability domain was assessed providing further guidelines for the in silico prediction purposes in decision support while performing risk assessment. QSAR models in the manuscript are available on-line through QsarDB repository for exploring and prediction services (http://hdl.handle.net/10967/106).

Environmental hazard of oil shale combustion fly ash.

The combined chemical and ecotoxicological characterization of oil shale combustion fly ash was performed. Ash was sampled from the most distant point of the ash-separation systems of the Balti and Eesti Thermal Power Plants in North-Eastern Estonia. The fly ash proved potentially hazardous for tested aquatic organisms and high alkalinity of the leachates (pH>10) is apparently the key factor determining its toxicity. The leachates were not genotoxic in the Ames assay. Also, the analysis showed that despite long-term intensive oil-shale combustion accompanied by considerable fly ash emissions has not led to significant soil contamination by hazardous trace elements in North-Eastern Estonia. Comparative study of the fly ash originating from the 'new' circulating fluidized bed (CFB) combustion technology and the 'old' pulverized-fired (PF) one showed that CFB fly ash was less toxic than PF fly ash. Thus, complete transfer to the 'new' technology will reduce (i) atmospheric emission of hazardous trace elements and (ii) fly ash toxicity to aquatic organisms as compared with the 'old' technology.

LuxCDABE--transformed constitutively bioluminescent Escherichia coli for toxicity screening: comparison with naturally luminous Vibrio fischeri.

We show that in vitro toxicity assay based on inhibition of the bioluminescence of recombinant Escherichia coli encoding thermostable luciferase from Photorhabdus luminescens is a versatile alternative to Vibrio fischeri Microtox™ test. Performance of two luxCDABE-transformed E. coli MC1061 constructs (pDNlux) and (pSLlux) otherwise identical, but having 100-fold different background luminescence was compared with the performance of V. fischeri. The microplate luminometer and a kinetic Flash-Assay test format was used that differently from Microtox test is also applicable for high throughput analysis. Toxic effects (30-s till 30-min EC(50)) of four heavy metals (Zn, Cd, Hg, Cu) and three organic chemicals (aniline, 3,5-dichloroaniline and 3,5-dichlorophenol) were studied. Both E. coli strains had comparable sensitivity and the respective 30-min EC(50) values highly correlated (log-log R(2) = 0.99; p < 0.01) showing that the sensitivity of the recombinant bacteria towards chemicals analyzed did not depend on the bioluminescence level of the recombinant cells. The most toxic chemical for all used bacterial strains (E. coli, V. fischeri) was mercury whereas the lowest EC(50) values for Hg (0.04-0.05 mg/L) and highest EC(50) values for aniline (1,300-1,700 mg/L) were observed for E. coli strains. Despite of that, toxicity results obtained with both E. coli strains (pSLlux and pDNlux) significantly correlated with V. fischeri results (log-log R(2) = 0.70/0.75; p < 0.05/0.01). The use of amino acids (0.25%) and glucose (0.05%)-supplemented M9 medium instead of leucine-supplemented saline significantly (p < 0.05) reduced the apparent toxicity of heavy metals to both E. coli strains up to three orders of magnitude, but had little or no complexing effect on organic compounds. Thus, P. luminescens luxCDABE-transformed E. coli strains can be successfully used for the acute toxicity screening of various types of organic chemicals and heavy metals and can replace V. fischeri in certain cases where the thermostability of luciferase >30 °C is crucial. The kinetic Flash Assay test format of the bioluminescence inhibition assay facilitates high throughput analysis. The assay medium, especially in case of testing heavy metals should be a compromise: optimal for the viability/luminescence of the recombinant test strain and of minimum complexing potential.

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).

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.

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.