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The intensive development of medical science has led to an increase in the availability and use of pharmaceutical products. However, nowadays, most of scientific attention has been paid to the native forms of pharmaceuticals, while the transformation products (TPs) of these substances, understood herein as metabolites, degradation products, and selected enantiomers, remain largely unexplored in terms of their characterization, presence, fate and effects within the natural environment. Therefore, the main aim of this study was to evaluate the toxicity of seven native compounds belonging to different therapeutic groups (non-steroidal anti-inflammatory drugs, opioid analgesics, beta-blockers, antibacterial and anti-epileptic drugs), along with the toxicity of their 13 most important TPs. For this purpose, an ecotoxicological test battery, consisting of five organisms of different biological organization was used. The obtained data shows that, in general, the toxicity of TPs to the tested organisms was similar or lower compared to their parent compounds. However, for example, significantly higher toxicity of the R form of ibuprofen to algae and duckweed, as well as a higher toxicity of the R form of naproxen to luminescent bacteria, was observed, proving that the risk associated with the presence of drug TPs in the environment should not be neglected. Electronic supplementary material The online version of this article (10.1007/s11356-020-08881-3) contains supplementary material, which is available to authorized users.

Wastewater from pharmaceutical and related industries contains many residual pharmaceutical components rich in color and high COD contents, which cannot be removed through the traditional wastewater treatment processes. Recently, microbial electrolysis ultraviolet cell (MEUC) process has shown its promising potential to remove recalcitrant organics because of its merits of wide pH range, iron-free, and without complications of iron sludge production. However, its application to the real pharmaceutical-rich industrial wastewater is still unknown. In this study, the MEUC process was validated with real ciprofloxacin-rich (6863.79 ± 2.21 µg L−1) industrial wastewater (6840 ± 110 mg L−1 of COD). The MEUC process achieved 100% removal of ciprofloxacin, 100% decolorization, and 99.1% removal of COD within 12, 60 and 30 h, respectively, when it was operated at pH-controlled at 7.8, applied voltage of 0.6 V, UV intensity of 10 mW cm−2, and cathodic aeration velocity of 0.005 mL min−1 mL−1. Moreover, fluorescence analysis showed that protein- and humic-like substances in such wastewater were effectively removed, providing further evidence of its high treatment efficiency. Furthermore, eco-toxicity testing with luminescent bacteria Vibro Feschri confirmed that the treated effluent was utterly non-toxic. The results demonstrated the broad application potential of MEUC technology for treating industrial wastewater.

Organic ligands are commonly believed to decrease the toxicity of metal ions, but there is few experimental evidence, especially for chromium (Cr(III)), which often coexists with organic compounds in industrial effluents. Here, the complexation of Cr(III) with acetate, lactate, l-tartrate, biphthalate and oxalate was tested under the conditions of a toxicity test, with high ion strength, by spectroscopic techniques. The stability constants of the complexes were found to follow the order Cr(III) oxalate > Cr(III) lactate > Cr(III) biphthalate > Cr(III) L-tartrate > Cr(III) acetate. Then, aquatic toxicity of Cr(III) to Photobacterium phosphoreum for a 15-min exposure period was tested in the absence and presence of organic ligands. Results unexpectedly show that the complexation of Cr(III) with acetic, lactate, l-tartrate and biphthalate resulted in enhanced toxicity to luminescent bacteria, whereas the coordination of Cr(III) with oxalate sharply alleviated the toxicity of individual oxalate and inorganic Cr(III), which was further confirmed by the scanning electron microscopy (SEM). Our findings show thus that organics do not always mitigate the toxicity of Cr(III) in acidic water.

It has been proven that the raw wastewater, secondary effluent and even reclaimed water may have toxic effects on aquatic organisms. In the present study, fractionation procedures combined with bioassays using luminescent bacteria were conducted to identify the fractions that contributed to the acute and long-term toxicity of municipal wastewater. Solid phase extraction was used to divide dissolved organic matter from the wastewater into three fractions, including non-polar, medium-polar and polar fraction. Among these fractions, although the acute toxicity of municipal wastewater was mainly caused by polar and medium-polar chemicals, the acute toxicity induced by the unit mass of the medium-polar fraction was the greatest. Using three kinds of resins, the organic substances in municipal wastewater were classified into six fractions, and the long-term toxicity of these fractions was further identified. The long-term toxicity of the hydrophobic neutrals, which were the primary toxic substances in raw wastewater, decreased after the conventional secondary biological treatment. Hydrophilic neutrals, which accounted for the majority of organic substances in the secondary effluent, were the main substances with long-term toxicity in the secondary effluent. The identification of fractions with acute and long-term toxicity in municipal wastewater is beneficial for further treatment to attenuate the ecotoxicity of wastewater before discharge into the aquatic environment.

This study evaluated the acute effects of nine different production chemicals typically employed in oil exploration on the toxicity of a synthetic produced water (PW). Bioassays with the Microtox® System were performed to monitor changes in the level of light emission of the marine luminescent bacteria Vibrio fischeri during exposure to the samples. The results show that synthetic PW is moderately toxic to these organisms, and the addition of oilfield chemicals significantly increases its toxicity. For most of the additives tested, the toxicity of the aqueous phase following partitioning against crude oil was not strongly altered by the presence of these chemicals. Synergistic effects occurred in the three different mixtures investigated. Among the additives studied, biocide, corrosion inhibitor, H2S scavenger, and surfactant were the most toxic for V. fischeri. Furthermore, the surfactant has been identified as the possible source of the acute toxicity observed.

In this paper, the releasing characteristics and biological toxicity of Tl, Hg, As and Sb in waste of Lanmuchang mercury-thallium mine were studied. The results indicated that strong acidity can significantly promote the release of Tl from waste. With the increase of pH, the release of Sb grew steadily, while Hg and As showed a trend of first increasing and then decreasing. Fe2(SO4)3 contributed less to the release of As and Sb than to that of Hg and Tl. FeCl3 significantly inhibited the release of As, Sb and Tl. In the leaching experiments of litter and root exudates, the lixiviums appeared neutral, and the litter and root exudates solution significantly reduced the release of Tl, and showed less toxicity to luminescent bacteria. However, they promoted the release of Hg, As and Sb at different levels.

Protecting the quality of lake watersheds by preventing and reducing their contamination is an effective approach to ensure the sustainability of the drinking water supply. In this study, acute toxicity assessment was conducted on the basis of acute bioluminescence inhibition assay using the marine bacterium Vibrio fischeri as the test organism and Luoma Lake drinking water source in East China as the research target. The suitable ranges of environmental factors, including pH value, organic matter, turbidity, hardness, and dissolved oxygen of water samples were evaluated for the toxicity testing of bioluminescent bacteria. The physicochemical characteristics of water samples at the selected 43 sites of Luoma Lake watershed were measured. Results showed that the variations in pH value (7.31–8.41), hardness (5–20 °d) and dissolved oxygen (4.44–11.03 mg/L) of Luoma Lake and its main inflow and outflow rivers had negligible impacts on the acute toxicity testing of V. fischeri. The luminescence inhibition rates ranged from −11.21% to 10.80% at the 43 sites. Pearson’s correlation analysis in the experiment revealed that temperature, pH value, hardness, and turbidity had no correlation with luminescence inhibition rate, whereas dissolved oxygen showed a weak statistically positive correlation with a Pearson correlation coefficient of 0.455 (p < 0.05).

Doxazosin (DOX), a selective alpha blocker, is widely used in medical therapy as an effective antihypertensive agent. It is a frequently prescribed drug and for this reason, environmental and ecotoxicological research is of great importance in terms of exposure and risk for both aquatic species and humans. In this study we focused on photolytic and TiO2 photocatalytic degradation processes of doxazosin under different simulated conditions, with the emphasis on identification of degradation products. Photolytic (without TiO2) experiments were performed in the presence and absence of oxygen, while photocatalytic degradation of doxazosin aqueous solution has been carried out under constant oxygen flow. DOX degradation was more efficient in the TiO2/UVA photocatalytic experiment than during photolytic processes (UVA and UVC, UVC-N2). LC-HRMS analyses with electrospray ionization allowed observing the formation of several major degradation products depending on the reaction conditions (presence or absence of oxygen, photocatalysis). The transformation products were identified based on exact mass measurements, isotopic distribution, and fragmentation pattern. Among them, dominated C17H21N5O3 and C17H23N5O4 (cleavage of the dioxane cycle), and C23H25N5O7 (hydroxylation). The detailed degradation pathway has been proposed. Toxicity testing with V. fischeri luminescent bacteria revealed higher toxicity of samples in photolytic rather than photocatalytic experiments which might be attributed to the formation of different products.

The production of biodegradable plastics is considered to be a way to reduce plastic waste issue. Among others, oxo-degradant additives enable a faster degradation of plastics in the environment. However, the introduction of these new materials could provoke the release of substances potentially toxic in the environment. This work determined and compared the toxicity of leachates from various additivated polymers (polyethylene, PE; polypropylene, PP; polystyrene, PS) upon different test organisms: plants (Sorghum saccharatum, Lepidium sativum, Sinapis alba, and Vicia faba), crustacean (Daphnia magna), and luminescent bacteria (Vibrio fischeri). Daphnia magna survival was mainly affected by PS and PP leachates (72% and 61% effect, respectively) while PS notably reduced the reproduction rate. On plants, only PP exerted a negative effect (S. saccharatum IG% 32.4), while V. fischeri always showed values around 50%. The data integration, through the Toxicity Test Battery Integrated Index (TBI) approach, allowed to rank the leachates toxicity as PE > PS > PP. This result could be mainly ascribable to the highest metals content in PE since no difference with organic compounds analysis was evidenced. In conclusion, since the polymers exerted dissimilar toxicity, the additive could not be considered the sole responsible of the measured toxicity, but its role in the enhancement of the virgin polymers leachates effects can be solidly hypothesized.

In this study, the applicability of UV absorbance at 254 nm (UV254) and volatile fatty acids (VFAs) to serve as reliable surrogates to predict acute toxicity of traditional Chinese medicine (TCM) wastewater was investigated. The medicine residues and VFAs were identified as main components of the TCM wastewater, and their individual and joint toxicity assays were operated with luminescent bacteria. The median effective concentration (EC50) values of medicine residues and VFAs were in the range of 26.46-165.55 mg/L and 11.45-20.58 g/L, respectively. The joint toxicity action modes of medicine residues, VFAs and medicine residues-VFAs were identified as additive, additive and synergistic respectively. UV254 and VFAs showed better correlations with acute toxicity according to the correlation analysis, compared with other conventional parameters. The regression model was a good fit for toxic unit (TU50) as a function of UV254 and VFAs according to the stepwise regression method (adjusted R2 = 0.836). Validation of the model to the pilot-scale samples provided satisfactory prediction results in the influent and hydrolysis acidification effluent samples tests, but for EGSB effluent and final effluent samples, the model needed further optimization. Surrogates prediction using UV254 and VFAs provided a valuable and cost-saving tool for rapid or on-line monitoring of acute toxicity of TCM wastewater.

Advanced oxidation processes (AOPs) have been widely applied for the advanced treatment of recalcitrant wastewater. However, the toxicity changes during AOPs are still unclear because in some cases the intermediate products could exhibit equivalent or even higher toxicity compared with the original pollutants. Therefore it is necessary to evaluate the toxicity evolution during AOPs. This review mainly focused on the toxicity changes of wastewater during various AOPs treatment, including Fenton/Fenton-like oxidation, ozonation/catalytic ozonation, persulfate radical-based oxidation, photocatalytic oxidation, ionizing radiation, electro-catalytic oxidation, ultrasound oxidation and wet oxidation. Firstly, the toxicity assessment methods were summarized, including acute toxicity (such as luminescent bacteria assay, fish assay, water flea assay and algae assay), genetic toxicity (Ames assay, SOS/umu assay, comet assay and micronucleus assay), estrogenic activity, immunity toxicity, and endocrine disrupting effect (such as yeast two-hybrid assay and fish endocrine disrupting assay), the applications of these methods in determining the toxicity changes of wastewater during AOPs treatment were summarized and analyzed in detail. Secondly, the toxicity prediction by computational simulation was introduced, mainly focusing on two common software, including TEST and ECOSAR. Thirdly, the strategies for eliminating the toxicity of wastewater after AOPs treatment were proposed. Finally, the concluding remarks and perspectives were presented for further study. This review will deepen to understand the toxicity evolution of wastewater during AOPs.

Highlights • UV-A irradiated epoxy coatings led to decreased ecotoxicological effects. • UV-A irradiated epoxy coatings released toxic transformation products. • 4-tert-Butylphenol was identified as main driver for ecotoxicological effects. • BPA and structural analogs contribute to estrogenic effects of irradiated materials., Organic coatings can guarantee long-term protection of steel structures due to causing a physical barrier against water and oxygen. Because of their mechanical properties and resistances to heat and chemicals, epoxy resin-based coatings are widely used for corrosion protection. Despite of the aromatic backbone and the resulting susceptibility to UV degradation, epoxy resins are frequently used as binding agent in top layers of anti-corrosion coating systems. Consequently, these organic polymers are directly exposed to sunlight and thus UV radiation. The present study was designed to investigate if toxic effects of epoxy resin-based-coatings are changed by UV-A irradiation. For this purpose, two epoxide-based top coatings were examined with and without UV aging for their bacterial toxicity and estrogenicity. In addition, chemical analyses were performed to identify released compounds as well as photolytic degradation products and to assign toxic effects to individual substances. UV-A irradiation of epoxy resin based top coatings resulted in an overall decrease of acute and specific ecotoxicological effects but as well to the formation of toxic transformation products. Both, in leachates of untreated and UV-A irradiated coatings, 4tBP was identified as the main driver of estrogenicity and toxicity to luminescent bacteria. BPA and structural analogs contributing to estrogenic effects in leachates were formed by UV-A irradiation. The combination of HPTLC coupled bioassays and LC-MS analyses supported the identification of bioactive compounds in terms of an effect-directed analysis. The present findings indicate that epoxide-based coatings are less suitable for the application as top coatings and more UV stable coatings like aliphatic polyurethanes should be preferred., Graphical abstract Image, graphical abstract

In the last decades, there has been an increasing concern about the human exposure to indoor dust. Therefore, it is imperative to assess the toxicity of indoor dust and associated dust extracts. In this study, the acute toxicity assessment of indoor dust was performed using a bioluminescence test, with Photobacterium phosphoreum T3 (PPT3) chosen as the test bacterium. The different indoor dust samples were collected from residences, offices, dormitories and laboratories in Shanghai, China. Our data reveal that PPT3 is more active to water-soluble ions and organic contaminants at low concentrations, while extract solutions elicit increased bacterial toxicity at high concentrations. The results of a bioluminescence assay by PPT3 indicated that the dust organic extracts exhibited increased toxicity compared with the water exacts. Dust extracts from the laboratory exhibited the greatest bacterial toxicity when compared with office, dormitory and residence samples. Moreover, office dust exhibited higher bacterial toxicity than residence dust. Furthermore, the comprehensive toxicity of dust on PPT3 was assessed by extracts toxicity -addition (i.e. IRaddition). The calculated values were close to the corresponding experimental data. The bioluminescence test showed the indoor dust samples are weakly toxic to PPT3, which are equivalent to 0.046–0.123 mg Hg•L−1. Different dust extracts among the different sampling sites showed varying toxicity to PPT3. This study provides some important information to understand the potential health risk from different indoor environment using a rapid bioluminescence assay.

Chlorination is essential to the safety of reclaimed water; however, this process leads to concern regarding the formation of disinfection byproducts (DBPs) and toxicity. This study reviewed the formation and control strategies for DBPs and toxicity in reclaimed water during chlorination. Both regulated and emerging DBPs have been frequently detected in reclaimed water during chlorination at a higher level than those in drinking water, indicating they pose a greater risk to humans. Luminescent bacteria and Daphnia magna acute toxicity, anti-estrogenic activity and cytotoxicity generally increased after chlorination because of the formation of DBPs. Genotoxicity by umu-test and estrogenic activity were decreased after chlorination because of destruction of toxic chemicals. During chlorination, water quality significantly impacted changes in toxicity. Ammonium tended to attenuate toxicity changes by reacting with chlorine to form chloramine, while bromide tended to aggravate toxicity changes by forming hypobromous acid. During pretreatment by ozonation and coagulation, disinfection byproduct formation potential (DBPFP) and toxicity formation potential (TFP) occasionally increase, which is accompanied by DOC removal; thus, the decrease of DOC was limited to indicate the decrease of DBPFP and TFP. It is more important to eliminate the key fraction of precursors such as hydrophobic acid and hydrophilic neutrals. During chlorination, toxicities can increase with the increasing chlorine dose and contact time. To control the excessive toxicity formation, a relatively low chlorine dose and short contact time were required. Quenching chlorine residual with reductive reagents also effectively abated the formation of toxic compounds.

The widespread use of organic ultraviolet (UV) filters in personal care products raises concerns about their potentially hazardous effects on human and ecosystem health. In this study, the toxicities of four commonly used benzophenones (BPs) UV filters including benzophenone (BP), 2-Hydroxybenzophenone (2HB), 2-Hydroxy-4-methoxybenzophenone (BP3), and 2-Hydroxy-4-methoxybenzophenone-5-sulfonicacid (BP4) in water were assayed in vitro using Vibrio fischeri, SOS/umu assay, and yeast estrogen screen (YES) assay, as well as in vivo using zebrafish larvae. The results showed that the luminescent bacteria toxicity, expressed as logEC50, increased with the lipophilicity (logKow) of BPs UV filters. Especially, since 2HB, BP3 and BP4 had different substituent groups, namely -OH, -OCH3 and -SO3H, respectively, these substituent functional groups had a major contribution to the lipophilicity and acute toxicity of these BPs. Similar tendency was observed for the genotoxicity, expressed as the value of induction ratio=1.5. Moreover, all the target BPs UV filters showed estrogenic activity, but no significant influences of lipophilicity on the estrogenicity were observed, with BP3 having the weakest estrogenic efficiency in vitro. Although BP3 displayed no noticeable adverse effects in any in vitro assays, multiple hormonal activities were observed in zebrafish larvae including estrogenicity, anti-estrogenicity and anti-androgenicity by regulating the expression of target genes. The results indicated potential hazardous effects of BPs UV filters and the importance of the combination of toxicological evaluation methods including in vitro and in vivo assays.

Phthalates has been widely used in children's toys as plastic plasticizers and softeners. The aim of this study was to extract chemically phthalates from 20 toy samples and determine toxicity using the Vibrio fischeri luminescent bacteria test. Subsequently the (the concentration producing 50% of maximal effect) was employed to characterize the toxicity of the sample and select the appropriate solvent from methanol, ethanol or glycerol in order to assess the adverse effects of di-n-butyl phthalate (DBP) or di-2-ethylhexyl phthalate (DEHP) and not solvent interference on Vibrio fischeri. Data showed that toxicity of glycerol was relatively low when the concentration was less than 0.44 mol/L. Therefore glycerol was selected as solvent. DBP and DEHP produced toxicity on Vibrio fischeri in the order of DEHP > DBP. The was 0.437 and 0.182 mol/L, respectively. This study provides the foundation for examination of combined toxic effects of the mixture of DBP and DEHP on Vibrio fischeri which is of signif...

Biological effects and ecotoxicity of NPs CeO2 with particle size Δ50 =16 nm and hydrobionts tolerance was measured in simulation based on test-reactions of a group of organisms representing major trophic levels of aquatic ecosystem. Biological activity of NPs CeO2 was noted against bacterial biosensor Ekolum, Сhlorella v. B.,Parameciumc., Daphniam. S. and Danior. Toxicity and hazard level of NPs CeO2 were measured using biotesting and hydrobionts. Toxicity of NPs CeO2 against luminescent bacteria Ekolum and Parameciumc. and Danior. was not established (L(E)С50 > 100 mg/l). In accordance with SGS and EC 93/67/EEC, NPs CeO2 is considered highly toxic (acute toxicity Level 1). Biomarkers and NPs CeO2 tolerant hydrobionts were determined. Range of tolerance of hydrobionts to NPs was established. The upper limit of tolerance was determined for certain hydrobionts by values of L(E)С50: 0.009 mg/l (by volume of Chl a) for Сhlorella v. B. and 46.15 mg/l (by mobility) for Daphniam. S.

As the application of nanoparticles (NPs) and their release to the environment has increased, it is important to verify their toxicity, with a special emphasis on particle solubilization and the interaction of NP mixtures. In the current study, a model luminescent bacteria, Vibrio fischeri, was employed to test the acute toxicity of individual NPs and their binary mixtures, including metal NPs (ZnNPs, CuNPs) and metal oxide NPs (ZnONPs, CuONPs). The independent action model was used to reflect the synergistic, additive, or antagonistic interactions of binary mixtures of these NPs. The results showed that the median effective concentration (EC50) inhibited the luminescence of V. fischeri were 20.5, 4.1, 11.6, and 118.7 mg L−1 for ZnNPs, CuNPs, ZnONPs, and CuONPs, respectively, suggesting that the toxicity of these NPs to V. fischeri were as the following order: CuNPs > ZnONPs > ZnNPs > CuONPs. The combined effect of NPs were found to be antagonistic for CuNPs-ZnONPs and CuNPs-CuONPs, synergistic for CuONPs-ZnNPs, CuNPs-ZnNPs, and ZnONPs-CuONPs, and additive for ZnNPs-ZnONPs, revealing a complex pattern of possible interactions. The differences of dissolved metal ions partly accounted for the different combined toxicity of binary mixtures of NPs. The findings have important implications for better understanding the true environmental risk of NP mixtures.

Often ingredients of personal care products are present in treated wastewaters, e. g grey water (GW), and are discharged into aquatic systems. Conventional treatment of GW does not fully eliminate micropollutants such as the UV filter substance 2-phenylbenzimidazole-5-sulfonic acid (PBSA). Photolysis has been proposed as an alternative treatment method for other micropollutants, but it is not clear yet whether it can also be used to eliminate PBSA. One goal of this study was to better understand the basic pathways involved in this process. It aimed to identify photo-transformation products (PTPs) by using, in the test conditions, an initial concentration of PBSA higher than those expected in the environment. The photolysis experiments were carried out using Xenon and UV lamps. Under Xenon irradiation only slight primary elimination was found. UV irradiation resulted in almost complete primary elimination of PBSA but not in full mineralization. Four isomeric mono-hydroxylated PTPs were identified by high resolution mass spectrometry (HRMS) which could be confirmed by other studies. A modified luminescent bacteria test (LBT) with Vibrio fischeri was employed to assess acute and chronic toxic effects of the irradiated photolytic mixtures. A strong correlation was found between the kinetics of two of the PTPs and luminescence inhibition indicating bacterial toxicity. Using a set of in silico quantitative structure-activity relationship (QSAR) models, this study also offered new insights concerning the environmental fate and toxicity of the TPs of PBSA as the TPs generated by UV-treatment are more persistent and partly more toxic than PBSA.

Predicting the crucial effect of single metal pollutants against the aquatic ecosystem has been highly debatable for decades. However, dealing with complex metal mixtures management in toxicological studies creates a challenge, as heavy metals may evoke greater toxicity on interactions with other constituents rather than individually low acting concentrations. Moreover, the toxicity mechanisms are different between short term and long term exposure of the metal toxicant. In this study, acute and chronic toxicity based on luminescence inhibition assay using newly isolated Photobacterium sp.NAA-MIE as the indicator are presented. Photobacterium sp.NAA-MIE was exposed to the mixture at a predetermined ratio of 1:1. TU (Toxicity Unit) and MTI (Mixture Toxic Index) approach presented the mixture toxicity of Hg2+ + Ag+, Hg2+ + Cu2+, Ag+ + Cu2+, Hg2+ + Ag+ + Cu2+, and Cd2+ + Cu2+ showed antagonistic effect over acute and chronic test. Binary mixture of Cu2+ + Zn2+ was observed to show additive effect at acute test and antagonistic effect at chronic test while mixture of Ni2+ + Zn2+ showing antagonistic effect during acute test and synergistic effect during chronic test. Thus, the strain is suitable and their use as bioassay to predict the risk assessment of heavy metal under acute toxicity without abandoning the advantage of chronic toxicity extrapolation.

Natural sunlight is a vital environmental element and plays a significant role in the ecological storage of reclaimed water (RW), but its impacts on RW quality are poorly understood. In this study, sunlight-induced changes in RW with a focus on dissolved organic matter (rDOM) and 52 residual micropollutants were investigated in the field during the summer and winter seasons. The results indicated that sunlight exposure led to the dissipation of chromophoric DOM (CDOM) in the summer (55% loss) and winter (19% loss) after 14 consecutive sunny days. During open storage of RW, CDOM absorption in UVC regions was preferentially removed in the summer, while during the winter there was preferential removal of CDOM in UVA regions. The results also showed higher fluorescent DOM (FDOM) removal in summer than in winter (49% and 28%, respectively). Results in both seasons indicated that humic acid-like compounds were the most photolabile fractions and were preferentially removed under sunlight exposure. Sunlight also induced attenuation of micropollutants in the summer and winter at reductions of 66% and 24% from the initial values, respectively. Significant attenuation (>75%) was only observed for endocrine-disrupting chemicals, pharmaceuticals, and sunscreens in the summer, but they accounted for 76% of the total concentrations. Vibrio fischeri toxicity tests demonstrated that sunlight constantly decreased the luminescent bacteria acute toxicity of RW, which was estimated to be caused mainly by the sunlight-induced changes of FDOM and CDOM, while the detected micropollutants could only explain 0.02%–2% of acute toxicity. These findings have important implications regarding our understanding of the ecological storage of reclaimed water and the contribution of management strategies.

Three distinct textile samples were investigated for color and toxicity (S1–chemical/textile industry; S2–final textile effluent; S3 - standard textile produced effluent–untreated blue). Radiation processing of these samples were carried out at Dynamitron Electron Beam Accelerator and color and toxicity removal were determined: color removal by radiation was 96% (40 kGy, S1); 55% (2.5 kGy, S2) and 90% (2.5 kGy, S3). Concerning toxicity assays, Vibrio fischeri luminescent bacteria demonstrated higher reduction after radiation than the other systems: removal efficiencies were 33% (20 kGy, S1); 55% (2.5 kGy, S2) and 33% (2.5 kGy, S3). Daphnia similis and Brachionus plicatilis fitted well for S3 effluents. Hard toxic volumes into biological treatment plant may be avoided if radiation would be previously applied in a real plant. Results reveled how indispensable is to run toxicity to more than one living-organism.

Advanced oxidation processes (AOPs) can degrade heavy metal complexes in wastewater to improve the removal efficiency of metals. However, the influences of AOP treatments on toxicity induced by metal complexes are not well understood. This study compared the toxicity induced by EDTA-copper (Cu) after UV/persulfate (PS) and UV/H2O2 treatments on luminescent bacteria and human HepG2 cells. The results showed that EDTA-Cu complexes decreased Cu toxicity in luminescent bacteria but increased the cytotoxicity in HepG2 cells, indicating species-specific toxicity. The UV/PS and UV/H2O2 treatments under most pH values and [oxidant]/[EDTA-Cu] conditions decreased the toxicity of EDTA-Cu in HepG2 cells but increased the toxicity in luminescent bacteria. When the ratio of [oxidant] to [EDTA-Cu] was 10, low toxicity in treated solutions was observed in both UV treatment processes. The alkaline precipitation treatment had a significant influence on toxicity reduction after UV/PS treatment; however, it had minimal influence on the UV/H2O2 treatment system. The Cu and total organic carbon (TOC) removal efficiency cannot completely explain the results of toxicity assays. EDTA-Cu intermediates might play important roles in changing the toxicity of EDTA-Cu after both UV treatments. This study provides insights into evaluating the treatment efficiency of UV/PS and UV/H2O2 on EDTA-Cu decomplexation.

Biotic ligand model (BLM) was extended to predict the toxicity of inorganic arsenate (iAs(V)) to the luminescent bacteria, Aliivibrio fischeri. As the pH increased from 5 to 9, the HAsO42- form predominated more than the H2AsO4- form did, and the EC50[As]T (50% effective iAs(V) concentration) decreased drastically from 3554 ± 393 to 39 ± 6 μM; thus, the HAsO42- form was more toxic to A. fischeri than H2AsO4-. As the HPO42- activity increased from 0 to 0.44 mM, the EC50{HAsO42-} values (50% effective HAsO42- activity) increased from 31 ± 6 to 859 ± 128 μM, indicating that the toxicity of iAs(V) decreased, owing to the competition caused by the structural similarity between iAs(V) and phosphate ions. However, activities of Ca2+, Mg2+, K+, SO42-, NO3-, and HCO3- did not significantly affect the EC50{HAsO42-} values. The BLM was reconstructed to take into account the effects of pH and phosphate, and the conditional binding constants for H2PO4-, HPO42-, H2AsO4-, and HAsO42- to the active binding sites of A. fischeri were obtained; 3.424 for logKXH2PO4, 4.588 for logKXHPO4, 3.067 for logKXH2AsO4, and 4.802 for logKXHAsO4. The fraction of active binding sites occupied by iAs(V) to induce 50% toxicity (fmix50%) was found to be 0.616.

In recent years, pharmaceuticals and personal care products (PPCPs) have emerged as significant pollutants of aquatic environments and have been detected at levels in the range of ng/L to μg/L. The source of PPCPs is humans and livestock that have been administered pharmaceuticals and subsequently excreted them via urine and feces. Unlike agricultural chemicals, the environmental dynamics of PPCPs is not examined and they would undergo structural transformation by environmental factors, e.g., sunlight, microorganisms and treatments in sewage treatment plants (STPs). Processing at STPs can remove various PPCPs; however, they are not removed completely and some persist in the effluents. In this study, we examined the degradation of 9 pharmaceuticals (acetaminophen, amiodarone, dapsone, dexamethasone, indomethacin, raloxifene, phenytoin, naproxen, and sulindac) by sunlight or UV, and investigated the ecotoxicological variation of degradation products. Sunlight (UVA and UVB) degraded most pharmaceuticals, except acetaminophen and phenytoin. Similar results were obtained with UVB and UVA. All the pharmaceuticals were photodegraded by UVC, which is used for sterilization in STPs. Ecotoxicity assay using the luminescent bacteria test (ISO11348) indicated that UVC irradiation increased the toxicity of acetaminophen and phenytoin significantly. The photodegraded product of acetaminophen was identified as 1-(2-amino-5-hydroxyphenyl)ethanone and that of phenytoin as benzophenone, and the authentic compounds showed high toxicity. Photodegraded products of PPCPs are a concern in ecotoxicology.

A novel tool, the DAMTA analyzer (Device for Analytical Monitoring and Toxicity Assessment), designed for fully automated toxicity measurements based on luminescent bacteria as well as for concomitant determination of chemical parameters, was developed and field-tested. The instrument is a robotic water analyzer equipped with a luminometer and a spectrophotometer, integrated on a thermostated reaction plate which contains a movable carousel with 80 cuvettes. Acute toxicity is measured on-line using a wild type Photobacterium phosphoreum strain with measurable bioluminescence and unaltered sensitivity to toxicants lasting up to ten days. The EC50 values of reference compounds tested were consistent with A. fischeri and P. phosphoreum international standards and comparable to previously published data. Concurrently, a laboratory trial demonstrated the feasibility of use of the analyzer for the determination of nutrients and metals in parallel to the toxicity measurements. In a prolonged test, the system was installed only in toxicity mode at the premises of the World Fair “Expo Milano-2015″, a high security site to ensure the quality of the supplied drinking water. The monitoring program lasted for six months during which ca. 2400 toxicity tests were carried out; the results indicated a mean non-toxic outcome of −5.5 ± 6.2%. In order to warrant the system's robustness in detecting toxic substances, Zn was measured daily with highly reproducible inhibition results, 70.8 ± 13.6%. These results assure that this novel toxicity monitor can be used as an early warning system for protection of drinking water sources from emergencies involving low probability/high impact contamination events in source water or treated water.

In this work, PM10 samples previously subjected to thorough chemical speciation and receptor modelling, have been investigated for their bio-toxicity using an inhibition test based on bacterial luminescence modulation when in contact with airborne particulate samples. The variation of light emission intensity from a luminescent bacteria strain, the Photobacterium phosphoreum, is proposed as an efficient proxy for the quantification of bio-toxic effects induced by airborne particulate matter. PM10 samples characterized by definite levels of pollutants from the pertaining air shed were found to induce a decrease in the bacterial bioluminescence intensity, expressed as percentage of Inhibition Ratio (IR%). This behaviour suggests the decay of this energy-consuming activity because of a toxic effect. Cluster analysis on chemical composition and IR% data provides evidence of a statistically significant association between the adverse effects on living cells and the range of specific chemical species in PM10.

Crassostrea virginica (the Eastern or American oyster) bioaccumulates pollutants from the water column, and therefore, its tissues can be used as bioindicators of past and present estuarine health. In this pilot project, we decided to investigate whether its tissues would be a suitable medium for toxicity testing using tissues from a variety of southern Texas locations of known and suspected anthropogenically impacted and unimpacted areas. We also conducted toxicity tests on sediments adjacent to oyster reefs using standard protocols for sediment toxicity. We tested the toxicity of tissues and sediments on the luminescent bacteria Vibrio fischeri, whose bioassays are commonly referred to by the trade name Microtox®. Microtox tests are quick, relatively inexpensive and sensitive to a range of contaminants. Evidence from this preliminary study suggests that conducting toxicity tests on oyster tissues may predict localized contamination better than when conducting toxicity tests on subtidal sediment. The refinement of these methods to use oyster tissues to detect contamination may be especially useful for environmental impact studies and/or studies where rapid and inexpensive information is needed.

The increasing application of nanoparticles (NPs) to a variety of new technologies has become a matter ofconcern due to the potential toxicity of these materials. Many questions about the fate of NPs in the environmentand the subsequent impact on ecosystems need to be answered. The aim of this work was to evaluate theecotoxicity of two alumina-based nanoceramics, γ-Al2O3 (NC) and Ni/ γ-Al2O3 (NiNC) by means of three differentstandardized tests: Biochemical Oxygen Demand (BOD5), bioassay with luminescent bacteria (Vibrio fischeri;Microtox), and bioassay on amphibian larvae (Rhinella arenarum) (AMPHITOX). BOD5 values of a verybiodegradable mixture (glucose/glutamic acid) decreased with the addition of NiNC(43.8%) and NC (31.6%)with respect to control samples (52.9%). Microtox test results indicated that NiNC presents higher toxicity thanNC, with EC50s values of 16.1% and 29.9% respectively; a reduced toxicity was observed, however, in presenceof organic matter, thus obtaining EC50s of 37.8% and 19.4%. The results of AMPHITOX test showed a significantincrease in the toxicity of both substances over time, the NiNC toxicity being greater than that of NC. The valuesof 96 h-LC50 and 504 h-LC50 determined for NiNC were 1.58 and 0.83 mg/L, respectively, and 14.5 and10.5 mg/L for NC samples. Amphibian larvae exhibited collapsed cavities, edema, axial flexures, and behavioralalterations as hyperkinesia and reduced movements. These results evidence the vulnerability of wildlife to xenobioticsand the need to develop specific standardized ecotoxicity tests in order to help environmental sustainabilityand natural species conservation. Fil: Svartz, Gabriela Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; Argentina Fil: Papa, Mariana Inés. Instituto Nacional de Tecnología Industrial; Argentina Fil: Gosatti, Marina. Comisión Nacional de Energía Atómica. Gerencia Complejo Tecnológico Pilcaniyeu; Argentina Fil: Jordán, Marianela Paula. Instituto Nacional de Tecnología Industrial; Argentina Fil: Soldati, Analía Leticia. Comisión Nacional de Energía Atómica. Gerencia Complejo Tecnológico Pilcaniyeu; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Samter, Paula. Instituto Nacional de Tecnología Industrial; Argentina Fil: Guraya, María M.. Comisión Nacional de Energía Atómica. Gerencia Complejo Tecnológico Pilcaniyeu; Argentina. Universidad Tecnológica Nacional. Facultad Regional Buenos Aires; Argentina Fil: Perez Coll, Cristina Silvia. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Perez Catán, Soledad. Comisión Nacional de Energía Atómica. Gerencia Complejo Tecnológico Pilcaniyeu; Argentina

Since aminophosphonate-based herbicides like glyphosate are currently one of the most popular and widely applied active agent in agrochemistry, there is an urgent need for searching new compounds among this family with potential herbicidal activity, but exhibiting low toxicity against surrounding environment. Six new (5-nitrofurfuryl)-derived aminophosphonates were synthesized for the first time and apart from the only one example of N-benzylamino(5-nitrofuryl)-methylphosphonic acid, it was the first time in the history, when this class of compounds was prepared. Their prospective and real biological properties have been followed up by evaluation of their preliminary ecotoxicology. They have been then investigated in aspect of their phytotoxicity against oat (A. sativa) and common radish (R. sativus) exhibiting moderate-to-severe toxicity for these plants. The significant selectivity towards radish (up to 3 times greater toxicity against radish) was observed in some cases. Title compounds were also tested in terms of their toxicity for freshwater crustaceans H. incongruens (ostracods) and marine luminescent bacteria A. fischeri. Although their harmful action on ostracods was not too much elevated, they were found to be highly toxic for bacteria. Various aspects of their ecotoxicity are discussed in this paper.

Slaughterhouse industry produces large volumes of polluted wastewater, which cause negative impacts on the environment. The objective of this study was to assess the effect of electron-beam irradiation on the ecotoxicity of slaughterhouse effluents with absorbed doses up to 35 kGy. Two acute toxicity assays were applied to evaluate the efficiency of irradiation onto toxicity of wastewater. The exposed living-organisms were a luminescent bacteria Vibrio fischeri, and a freshwater microcrustacean Daphnia similis. Also, the total organic carbon was analysed in order to determine any possible organic carbon removal after irradiation. The ecotoxicological results evidenced that both living-organisms were suitable for the measurements. Therefore, the results demonstrated the toxicity of the effluent and its similarity for both organisms as well as the potential of radiation to reduce these effects. The 35 kGy dose was very effective for reducing toxic effects of slaughterhouse wastewater for daphnids suggesting that ionizing radiation could be used as a tool for removing toxic charge of such effluents. The type of contamination presented by the effluent justify the needs for alternatives of treatment.

The purpose of this study was to discuss the effect of the chemical structure of anionic and non-ionic surfactants and surface activity on toxicity. Single and binary mixtures of three ether carboxylic derivative sur- factants and three amine-oxide-based non-ionic surfactants were used. Toxicity was determined using three test organisms: freshwater crustaceans (Daphnia magna), luminescent bacteria (Vibrio fischeri), and microalgae (Selenastrum capricornutum). The toxicity of surfactants is related to the hydrophobic alkyl chain, the degree of eth- oxylation, and the critical micelle concentration of sur- factants. Relationships found agreed with the fact that the lower toxicity is shown by the shorter alkyl chain. There is a strong relation between surface activity and toxicity: the toxicity increased as the CMC of the surfactant or mixtures of surfactants decreased. Commercial products are formu- lated using surfactants mixtures, so it is important to know their behavior using an easily measured property: the least toxic mixtures were formed by the surfactants having lower individual toxicity. Around the CMC, our data show a synergism for the binary mixtures. The results have given rise to a classification of the different surfactants and their mixtures according to the organism test, as safe, harmful or toxic. V. fischeri was in general the most sensitive micro- organism to the toxic effect of the surfactants, followed by Daphnia magna, while Selenastrum capricornutum was more tolerant. These results can be useful for selecting technically efficient surfactants and their mixtures with a lower ecotoxicity on the aquatic environment.

Following urbanization, a large number of urban rivers were contaminated and turned to black-odor rivers. The traditional approach for detecting water quality is based on chemical or physical analysis. However, biological toxicity of black-odor water has been less addressed. As two typical black-odor rivers, Jiushanwai River (JS) and Shanxia River (SX) are tributaries of Wen-Rui Tang River in Wenzhou (south of China). The eco-safety of the urban rivers was evaluated by bioassay for water toxicity in this study. Ten and 5 sampling sites were respectively set along JS and SX. Water samples were collected monthly from October 2010 to October 2011. The general physical and chemical parameters of river water were monitored. In order to investigate the ecotoxicological effects of black-odor water, the following bioassays were used: (1) Fish acute toxicity test (Danio rerio, comprehensive toxicity), (2) luminescent bacteria bioassay (Qinghaiensis vibrio, toxicity to bacteria), and (3) tropical claw embryo assay (Xenopus tropicalis, embryo toxicity). Biotoxicity of black-odor rivers water was demonstrated by D. rerio, Q. vibrio, and X. tropicalis embryos. Toxicological effects of black-odor water were respectively shown by mortality of zebrafish, and by the relative inhibitory light rate of luminescent bacteria. However, luminescent bacteria were more sensitive to inspect biotoxicity than zebrafish. In X. tropicalis embryos test, toxicological effects of black-odor water were mostly shown by embryos’ survival rate and teratogenic rate. Bioassay results showed that toxicity of SX water was higher than that of JS water, especially in summer. Statistical analysis of luminescent bacteria toxicity test showed that biotoxicity of SX and JS was high in summer, but low in winter and spring. The seasonal changes of water toxicity of the black-odor river were positively correlative with changes of water temperature (p

Tetracycline was exposed to ozone by bubbling at different pH levels, and the acute toxicity of ozonation byproducts was tested by luminescent bacteria (Vibrio qinghaiensis sp.-Q67). Residual tetracycline in the solutions was detected by high performance liquid chromatography (HPLC). The results indicated that tetracycline was immediately degraded by ozonation and the degradation was favored at higher pH. Tetracycline (20 mg L−1) was completely degraded after 5 min of ozonation. However, total organic carbon (TOC) hardly decreased even though the ozonation time was extended to 40 min, suggesting that the ozonation process could not mineralize tetracycline. The luminescent bacteria assay indicated that the toxicity of the tetracycline solution was changed after ozonation. Chemical oxygen demand (COD) dropped by 15%, and the BOD5/COD ratio increased from 0·09 to 0·26, suggesting that the biodegradability of tetracycline was enhanced after 20 min of ozonation treatment.

Luminescent bacterial test is a fast and sensitive method for acute toxicity assessment of water and wastewater. In this study, an improved toxicity testing method was developed using the freshwater luminescent bacteria Vibrio qinghaiensis sp.-Q67 that involved pretreatment of water samples with reverse osmosis (RO) to eliminate the interferences caused by nutrients in concentrated samples and to improve the reliability and sensitivity of the analysis. Because water samples contain low concentrations of several target toxic substances, rapid acute toxicity testing method that is commonly employed does not achieve enough sensitivity. The proposed RO pretreatment could effectively enrich organic and inorganic substances in water samples to enable a more effective and sensitive toxicity evaluation. The kinetic characteristics of toxicity of raw sewage and secondary effluent were evaluated based on the relative luminescence unit (RLU) curves and time-concentration-effect surfaces. It was observed that when the exposure time was prolonged to 8-h or longer, the bacteria reached the logarithmic growth stage. Hence, the stimulating effects of the coexisting ions (such as Na+, K+, NO3-) in the concentrated samples could be well eliminated. A 10-h exposure time in proposed Q67 test was found to quantitatively evaluate the toxicity of the organic and inorganic pollutants in the RO-concentrated samples. © 2013 Elsevier Inc.

Triclosan, one of the most widely used disinfectants, has been found to be toxic to animals and human beings. In this paper, triclosan was degraded on TiO 2 nanotube arrays, using a photoelectrocatalytic (PEC) process under UV illumination, with Na 2 SO 4 as the supporting electrolyte. The effect of bias potential was investigated and the results showed that 0 V was the most appropriate potential for the degradation of triclosan. In 30 min, 78.7% of triclosan had degraded during the PEC process. Intermediate analysis showed that 2,7-dichlorodibenzodioxin (DCDD) had formed during the degradation. The toxicity change during the PEC process was investigated using a luminescent bacteria test, with the results demonstrating that the toxicity of the reaction liquid decreased at the beginning and subsequently increased to a stable level. The indications were that some intermediates such as 2,7-dichlorodibenzodioxin was more toxic and stable than triclosan in the solution.

The present study demonstrates development of a rapid testing protocol based on a small portable luminometer using flash kinetic assessment of bacterial bioluminescence. The laboratory comparisons based on six model organic toxicants and two metals showed significant correlations between responses of freshwater bacteria Photorhabdus luminescens and standard marine bacterial species Vibrio fisheri. While P. luminescens was less sensitive in standard arrangements, the responses of both organisms were comparable in the newly introduced portable luminometer setup. The applicability and reproducibility of the portable luminometer protocol was further demonstrated in the assessment of 43 European wastewater effluents that were simultaneously tested for toxicity and analysed for 150 organic and 20 inorganic contaminants grouped into 13 major chemical classes. Clear association between the toxic responses in both compared bacterial species and the elevated levels of inorganic compounds (toxic metals), chlorophenols and benzotriazole anticorrosives was observed. The new protocol with a portable luminometer provides a fast (30 s) response and may be used as a tool for rapid in situ toxicity evaluation of freshwater environmental samples such as effluents.

Drinking water treatment residue (DWTR) seems to be very promising for controlling lake sediment pollution. Logically, acquisition of the potential toxicity of DWTR will be beneficial for its applications. In this study, the toxicity of DWTR and sediments amended with DWTR to Aliivibrio fischeri was evaluated based on the Microtox(®) solid and leachate phase assays, in combination with flow cytometry analyses and the kinetic luminescent bacteria test. The results showed that both solid particles and aqueous/organic extracts of DWTR exhibited no toxicity to the bacterial luminescence and growth. The solid particles of DWTR even promoted bacterial luminescence, possibly because DWTR particles could act as a microbial carrier and provide nutrients for bacteria growth. Bacterial toxicity (either luminescence or growth) was observed from the solid phase and aqueous/organic extracts of sediments with or without DWTR addition. Further analysis showed that the solid phase toxicity was determined to be related mainly to the fixation of bacteria to fine particles and/or organic matter, and all of the observed inhibition resulting from aqueous/organic extracts was identified as non-significant. Moreover, DWTR addition not only had no adverse effect on the aqueous/organic extract toxicity of the sediment but also reduced the solid phase toxicity of the sediment. Overall, in practical application, the solid particles, the water-soluble substances transferred to surface water or the organic substances in DWTR had no toxicity or any delayed effect on bacteria in lakes, and DWTR can therefore be considered as a non-hazardous material.

This study was conducted to assess the microbial toxicity of ionic silver solution () and silver nanoparticle suspension () based on the Microtox bioassay. In this test, the light inhibition of luminescent bacteria was measured after 15 and 30 min exposure to aqueous solutions and soils spiked with a dilution series of and . The resulting dose-response curves were used to derive effective concentration (EC25, , EC75) and effective dose (, , ) that caused a 25, 50 and 75% inhibition of luminescence. In aqueous solutions, value of after 15 min exposure was determined to be and remarkably lower than value of with . This revealed that was more toxic to luminescent bacteria than . In soil extracts, however, value of with 196 mg kg-1 was higher than value of with , indicating less toxicity of in soils. The reduced toxicity of in soils can be attributed to a partial adsorption of ionic on soil colloids and humic acid as well as a partial formation of insoluble AgCl with NaCl of Microtox diluent. This resulted in lower concentration of active Ag in soil extracts obtained after 1 hour shaking with than that spiked with . With longer exposure time, EC and ED values of both and decreased, so their toxicity increased. The toxic characteristics of silver nanomaterials were different depending on existing form of Ag (, ), reaction medium (aqueous solution, soil), and exposure time.

This study was focused on photodegradation of acetaminophen (AA) exposed to ultraviolet (UV) irradiation at 254 nm delivered by a system similar to that used for sterilization in sewage treatment plants. The degradation of AA during irradiation for up to 96 h was monitored by means of high-performance liquid chromatography (HPLC). Based on the mass/charge ratio ( m / z ) of the protonated ion and the mass fragmentation pattern in electrospray ionization time-of-flight mass spectrometry (ESI-TOF/MS/MS), the photoproduct was identified as 1-(2-amino-5-hydroxyphenyl)ethanone ( 1 ). Examination of toxicity by means of a luminescent bacteria test (ISO11348) indicated that AA solution was nontoxic, whereas photo-exposed AA solution was toxic (EC 50 : 29.46 mg/L). The toxicity of synthetic compound 1 was shown to account for a substantial part of the toxicity of photo-exposed AA. These results indicate the importance of investigating not only parent compounds, but also photoproducts the risk assessment of pharmaceuticals in aquatic environments.

The fate of canola oil in water under aerobic conditions was studied in respirometry tests. First-order biodegradation rates were estimated for a refined oil (commercial canola oil, CCO), 0.0037 h−1, and for two synthetic oils, one prepared with mono-acid triacylglycerols (TAO) 0.0048 h−1, and the other with free fatty acids and glycerol (FAO) 0.0038 h−1. Two abiotic factors limited substrate bioavailability: the autoxidation of the oleoyl chains and the insolubility of fully saturated triacylglycerols. In addition, the ecotoxic impact of the oils was assessed by Microtox® toxicity. For the acylglycerol oils, luminescent bacteria inhibition was not detected in the water phase, whereas solid-phase EC50 values below 1.8% sample volume were measured. Toxicity was observed in the aqueous phase (EC50