Sulfamethoxazole is Metabolized and Mineralized at Extremely Low Concentrations.

The presence of organic micropollutants in water and sediments motivates investigation of their biotransformation at environmentally low concentrations, usually in the range of µg L-1. Many are biotransformed by cometabolic mechanisms; however, there is scarce information concerning their direct metabolization in this concentration range. Threshold concentrations for microbial assimilation have been reported in both pure and mixed cultures from different origins. The literature suggests a range value for bacterial growth of 1-100 µg L-1 for isolated aerobic heterotrophs in the presence of a single substrate. We aimed to investigate, as a model case, the threshold level for sulfamethoxazole (SMX) metabolization in pure cultures of Microbacterium strain BR1. Previous research with this strain has covered the milligram L-1 range. In this study, acclimated cultures were exposed to concentrations from 0.1 to 25 µg L-1 of 14C-labeled SMX, and the 14C-CO2 produced was trapped and quantified over 24 h. Interestingly, SMX removal was rapid, with 98% removed within 2 h. In contrast, mineralization was slower, with a consistent percentage of 60.0 ± 0.7% found at all concentrations. Mineralization rates increased with rising concentrations. Therefore, this study shows that bacteria are capable of the direct metabolization of organic micropollutants at extremely low concentrations (sub µg L-1).

Extrapolation of cytotoxic masked effects in planar in vitro assays.

The masking of specific effects in in vitro assays by cytotoxicity is a commonly known phenomenon. This may result in a partial or complete loss of effect signals. For common in vitro assays, approaches for identifying and quantifying cytotoxic masking are partly available. However, a quantification of cytotoxicity-affected signals is not possible. As an alternative, planar bioassays that combine high-performance thin layer chromatography with in vitro assays, such as the planar yeast estrogen screen (p-YES), might allow for a quantification of cytotoxically affected signals. Affected signals form a typical ring structure with a supressed or completely lacking centre that results in a double peak chromatogram. This study investigates whether these double peaks can be used for fitting a peak function to extrapolate the theoretical, unaffected signals. The precision of the modelling was evaluated for four individual peak functions, using 42 ideal, undistorted peaks from estrogenic model compounds in the p-YES. Modelled ED50-values from bisphenol A (BPA) experiments with cytotoxically disturbed signals were 13 times higher than for the apparent data without compensation for cytotoxicity (320 ± 63 ng versus 24 ± 17 ng). This finding has a high relevance for the modelling of mixture effects according to concentration addition that requires unaffected, complete dose-response relationships. Finally, we applied the approach to results of a p-YES assay on leachate samples of an elastomer material used in water engineering. In summary, the fitting approach enables the quantitative evaluation of cytotoxically affected signals in planar in vitro assays and also has applications for other fields of chemical analysis like distorted chromatography signals.

JRODOS customization for Pakistan and assessment of the consequences of a hypothetical nuclear accident.

Since the Chernobyl disaster in 1986, decision support systems and modelling tools have been utilized in response to nuclear and radiological emergencies. The java-based real-time online decision support system (JRODOS) is a decision support tool that can be utilised in response to an emergency in managing off-site radiological consequences. This article documents the customization and use of JRODOS for Pakistan. JRODOS was tailored to the local Pakistan conditions, and a case study of a theoretical nuclear power plant accident was used to assess JRODOS's feasibility as a decision support tool. A worst-case probabilistic accident scenario was used to identify zones and areas where urgent protective actions, early protective actions and food restriction and other response actions could be required. The areas and distances identified for the implementation of protective and response actions for such a hypothetical accident were found to be in agreement with the emergency planning zones and distances suggested by the International Atomic Energy Agency (IAEA). Additionally, the implications of meteorological and source term input parameters on predicting the radiation doses to members of the public were investigated. It was identified that the output of such tools strongly depends on the availability and accuracy of the input parameters, such as radioactive release and meteorological data. Limitations and uncertainties associated with these tools need to be considered in deciding on protective and other response actions in response to a nuclear accident. As established by the IAEA, protective and other response actions need to be applied on a graded approach, taking into account the protection strategy and uncertainties and limitations in the available information and criteria, based on the conditions at the facility and off-site.

Passive Sampling and Dosing of Aquatic Organic Contaminant Mixtures for Ecotoxicological Analyses.

Toxicity results from exposure to mixtures of organic contaminants. Assessing this using ecotoxicity bioassays involves sampling of the environmental mixture and then introducing this into the test. The first step is accounting for the bioavailable levels of all mixture constituents. Passive sampling specifically targets these bioavailable fractions but the sampler-accumulated mixture varies with the compound and sampler properties as well as time. The second step involves reproducing and maintaining the sampled mixture constituents in the bioassay. Passive sampler extraction and spiking always leads to a skewed mixture profile in the test. Alternatively, the recovered passive samplers might be directly used in passive dosing mode. Here, the reproduced contaminant mixture depends on whether kinetic or equilibrium sampling applies. These concepts were tested for determining the combined toxicity of laboratory and field mixtures of aquatic contaminants in the Microtox and ER-Calux bioassays. Aqueous sample extraction and spiking, passive sampler extraction and spiking, and passive sampling and dosing were compared for first sampling and then introducing mixtures in toxicity bioassays. The analytical and toxicity results show that the correct way to first sample the bioavailable mixture profile, and then to reproduce and maintain this in the toxicity test, is by combining equilibrium passive sampling and dosing.

Kinetic Passive Sampling: In Situ Calibration Using the Contaminant Mass Measured in Parallel Samplers with Different Thicknesses.

The use of single-phase passive samplers is a common method for sampling bioavailable concentrations of hydrophobic aquatic pollutants. Often such samplers are used in the kinetic stage, and in situ calibration is necessary. Most commonly, exchange kinetics are derived from the release rates of performance reference compounds (PRCs). In this study, a complementary calibration approach was developed, in which measuring the contaminant mass ratio (CMR) from two samplers with different thicknesses allows the dissolved concentrations to be determined. This new CMR calibration was tested (1) in a laboratory experiment with defined and constant concentrations and (2) in the field, at a storm water retention site. Silicone passive samplers with different thicknesses were used to sample a range of dissolved polycyclic aromatic hydrocarbons. In the laboratory study, the concentrations derived from the CMR calibration were compared with those from water extraction and passive dosing and differences below a factor 2 were found. In the field study, CMR-derived concentrations were compared to those from PRC calibration. Here, differences ranged by only a factor 1 to 3 between both methods. These findings indicate that the CMR calibration can be applied as a stand-alone or complementary calibration method for kinetic passive sampling.

Bioactivation of Quinolines in a Recombinant Estrogen Receptor Transactivation Assay Is Catalyzed by N-Methyltransferases.

Hydroxylation of polyaromatic compounds through cytochromes P450 (CYPs) is known to result in potentially estrogenic transformation products. Recently, there has been an increasing awareness of the importance of alternative pathways such as aldehyde oxidases (AOX) or N-methyltransferases (NMT) in bioactivation of small molecules, particularly N-heterocycles. Therefore, this study investigated the biotransformation and activity of methylated quinolines, a class of environmentally relevant N-heterocycles that are no native ligands of the estrogen receptor (ER), in the estrogen-responsive cell line ERα CALUX. We found that this widely used cell line overexpresses AOXs and NMTs while having low expression of CYP enzymes. Exposure of ERα CALUX cells to quinolines resulted in estrogenic effects, which could be mitigated using an inhibitor of AOX/NMTs. No such mitigation occurred after coexposure to a CYP1A inhibitor. A number of N-methylated but no hydroxylated transformation products were detected using liquid chromatography-mass spectrometry, which indicated that biotransformations to estrogenic metabolites were likely catalyzed by NMTs. Compared to the natural ER ligand 17ß-estradiol, the products formed during the metabolization of quinolines were weak to moderate agonists of the human ERα. Our findings have potential implications for the risk assessment of these compounds and indicate that care must be taken when using in vitro estrogenicity assays, for example, ERα CALUX, for the characterization of N-heterocycles or environmental samples that may contain them.

Directed OmniChange Evolution Converts P450 BM3 into an Alkyltrimethylammonium Hydroxylase.

Cetyl-trimethylammonium bromide (CTAB) is a widely used cationic surfactant that is biodegradable in nature. CTAB biodegradation requires hydroxylation in the first step, which is rate-limiting and crucial for solubility in water. In this study, the OmniChange multi-site mutagenesis method was applied to reengineer the P450 BM3 substrate specificity towards the hydroxylation of CTAB by simultaneous mutagenesis of four previously reported positions (R47, Y51, F87, and L188). 1740 clones from the P450 BM3 OmniChange library were screened with the NADPH depletion assay. A total of 696 clones were rescreened with the NADPH depletion and an Ampliflu™ Red/ horseradish peroxidase based H2 O2 detection assay. Several improved P450 BM3 variants were identified and finally four were kinetically characterized with respect to CTAB hydroxylation, based on both performance and coupling efficiency. Based on NADPH consumption, the P450 BM3 variant P3A8 (R47E/Y51M/F87V/L188E) displayed an initial activity (64.9±4.8 s-1 , 13.5-fold increased activity compared with wild-type P450 BM3), which nearly matches the specific activity for its natural fatty acid substrate (palmitic acid (32-122 s-1 )). Variant P3A8 showed high coupling efficiency (92.5 %), whereas wild-type P450 BM3 displayed a low coupling efficiency (0.5 %). HPLC-MS/MS detection confirmed that P3A8 and P2E7 (R47D/Y51L/F87V/L188A) form 13 and 35 times more 2-hydroxylated CTAB than P450 BM3. In addition, di-hydroxylated CTAB products were detected for all four investigated P450 BM3 variants (up to a yield of 77 %; P3A8). Di-hydroxylated quaternary amines are highly interesting bolaform surfactants with a high hydrophilicity (surface contact angle: θ=16.7°).

Passive dosing of polycyclic aromatic hydrocarbon (PAH) mixtures to terrestrial springtails: linking mixture toxicity to chemical activities, equilibrium lipid concentrations, and toxic units.

A 7-day mixture toxicity experiment with the terrestrial springtail Folsomia candida was conducted, and the effects were linked to three different mixture exposure parameters. Passive dosing from silicone was applied to tightly control exposure levels and compositions of 12 mixture treatments, containing the polycyclic aromatic hydrocarbons (PAHs) naphthalene, phenanthrene, and pyrene. Springtail lethality was then linked to sum chemical activities (∑a), sum equilibrium lipid concentrations (∑C(lipid eq.)), and sum toxic units (∑TU). In each case, the effects of all 12 mixture treatments could be fitted to one sigmoidal exposure-response relationship. The effective lethal chemical activity (La50) of 0.027 was well within the expected range for baseline toxicity of 0.01-0.1. Linking the effects to the lipid-based exposure parameter yielded an effective lethal concentration (LC(lipid eq 50)) of 133 mmol kg(-1) lipid in good correspondence with the lethal membrane burden for baseline toxicity (40-160 mmol kg(-1) lipid). Finally, the effective lethal toxic unit (LTU50) of 1.20 was rather close to the expected value of 1. Altogether, passive dosing provided tightly controlled mixture exposure in terms of both level and composition, while ∑a, ∑C(lipid eq.), and ∑TU allowed baseline toxicity to be linked to mixture exposure.

Baseline toxic mixtures of non-toxic chemicals: "solubility addition" increases exposure for solid hydrophobic chemicals.

This study addresses the question whether hydrophobic organic chemicals exerting no toxicity at their solubility limit (saturation) can form a toxic mixture. Spiking methods generally do not allow testing exactly at saturation without introducing microcrystals. Passive dosing was thus applied to test the acute toxicity of several high melting point PAHs and their mixtures at the respective saturation levels to aquatic and terrestrial invertebrates. With the aquatic Daphnia magna, anthracene, chrysene, and benzo(a)pyrene resulted in no or limited acute toxicity (0-20%), whereas binary and tertiary mixtures of these resulted in significant acute toxicity (70-88%). Toxicity of PAHs and their mixtures could be fitted with one (sum) chemical activity-response curve in accordance with a similar mode of toxic action (i.e., concentration addition). The effective chemical activity (Ea-50) of 0.029 and the effective concentration on a lipid basis (EC(lipid, eq.)-50) of 95.7 mM were well within the range for baseline toxicity. Similar mixtures showed less toxicity to the terrestrial Folsomia candida due to steady-state body-burdens being below equilibrium partitioning levels. The results of the present study raise questions about the focus of risk assessment schemes and toxicity testing guidelines on individual substances, since apparently non-toxic chemicals might become toxic in a mixture.

The dosing determines mutagenicity of hydrophobic compounds in the Ames II assay with metabolic transformation: passive dosing versus solvent spiking.

The Ames II bacterial mutagenicity assay is a new version of the standard Ames test for screening chemicals for genotoxic activity. However, the use of plastic micro-titer plates has drawbacks in the case of testing hydrophobic mutagens, since sorptive and other losses make it difficult to control and define the exposure concentrations, and they reduce availability for bacterial uptake or to the S9 enzymes. With passive dosing, a biocompatible polymer such as silicone is loaded with the test compound and acts as a partitioning source. It compensates for any losses and results in stable freely dissolved concentrations. Passive dosing using silicone O-rings was applied in the Ames II assay to measure PAH mutagenicity in strains TA98 and TAMix - a mixture of six different bacterial strains detecting six different base-pair substitutions - after metabolic activation by S9. Initially, 10 PAHs were tested with passive dosing from saturated O-rings, aiming at levels in the test medium close to aqueous solubility. Fluoranthene, pyrene and benzo(a)pyrene were mutagenic in both TA98 and TAMix, whereas benz(a)anthracene was mutagenic in TA98 only. The concentration-dependent mutagenic activity of benzo(a)pyrene was then compared for passive dosing and solvent spiking. With spiking, nominal concentrations greatly exceeded aqueous solubility before mutagenicity was observed, due to sorptive losses and limiting dissolution kinetics. In contrast, the passive dosing concentration-response curves were more reproducible, and shifted towards lower concentrations by several orders of magnitude. This study raises fundamental questions about how to introduce hydrophobic test substances in the Ames II assay with biotransformation, since the measured mutagenicity not only depends on the compound potency but also on its supply, sorption and consumption during the assay.

Fermented biochar has a markedly different effect on fate of pesticides in soil than compost, straw, and a mixed biochar-product.

Current knowledge about how biochars affect the fate of pesticides in soil is based on studies that used pure biochars. After finding that an additional biological post-pyrolysis treatment, such as co-composting or lactic fermentation, is required for biochars for superior performance in temperate arable soils, a knowledge gap formed of how such further processed biochar products would affect the fate of pesticides in soil. This study compared the effects of a novel fermented biochar alone or mixed with biogas residues on the fate of two pesticides, 4-chloro-2-methylphenoxyacetic acid (MCPA) and metalaxyl-M, in a temperate arable soil to the traditional organic amendments wheat straw and compost. The fate of 14C-labeled MCPA was markedly affected in different ways. Fermented biochar effectively reduced the water-extractability and mineralization due to adsorption that was comparable to adsorption strengths reported for pure biochars. However, this effect was weak for the biochar mixed with biogas residues. Straw reduced water-extractable amounts due to increased biodegradation and formation of likely biogenic non-extractable residues of MCPA. In contrast, compost decelerated mineralization and increased the water solubility of the MCPA residues due to released dissolved organic matter. The amendments' effects were minor regarding 14C-metalaxyl-M, except for the fermented biochar which again reduced water-extractability and delayed degradation due to adsorption. Thus, the effects of the organic amendments differed for the two pesticide compounds with only the fermented biochar's effect being similar for both. However, this effect was no longer present in the mixed product containing 20% biochar. Our findings clearly show that biologically treated biochar-containing products can affect the fate of pesticides in soil very differently, also when compared to traditional organic amendments. Such impacts and their desirable and undesirable ecotoxicological implications need to be considered before the large-scale application of biochars to temperate arable soils.

Measuring binding and speciation of hydrophobic organic chemicals at controlled freely dissolved concentrations and without phase separation.

The binding and speciation of hydrophobic organic chemicals (HOCs) in aqueous solutions were determined by controlling chemical activity and measuring total concentrations. Passive dosing was applied to control chemical activities of HOCs in aqueous solutions by equilibrium partitioning from a poly(dimethylsiloxane) polymer preloaded with the chemicals. The HOC concentrations in the equilibrated solutions [C(solution(eq))] and water [C(water(eq))] were then measured. Free fractions of the HOCs were determined as C(water(eq))/C(solution(eq)), whereas enhanced capacities (E) of the solutions for HOCs were determined as C(solution(eq))/C(water(eq)). A mixture of polycyclic aromatic hydrocarbons served as model analytes, while humic acid, sodium dodecyl sulfate, hydroxypropyl-ß-cyclodextrin, and NaCl served as model medium constituents. The enhanced capacities were plotted versus the concentrations of medium constituents, and simple linear regression provided precise partition ratios, salting out constants, and critical micelle concentrations. These parameters were generally in good agreement with published values obtained by solid phase microextraction and fluorescence quenching. The very good precision was indicated by the low relative standard errors for the partition ratios of 0.5-8%, equivalent to 0.002-0.03 log unit. This passive dosing approach allows binding and speciation of HOCs to be studied without any phase separation steps or mass balance assumptions.

Dynamic passive dosing for studying the biotransformation of hydrophobic organic chemicals: microbial degradation as an example.

Biotransformation plays a key role in hydrophobic organic compound (HOC) fate, and understanding kinetics as a function of (bio)availability is critical for elucidating persistence, accumulation, and toxicity. Biotransformation mainly occurs in an aqueous environment, posing technical challenges for producing kinetic data because of low HOC solubilities and sorptive losses. To overcome these, a new experimental approach based on passive dosing is presented. This avoids using cosolvent for introducing the HOC substrate, buffers substrate depletion so biotransformation is measured within a narrow and defined dissolved concentration range, and enables high compound turnover even at low concentrations to simplify end point measurement. As a case study, the biodegradation kinetics of two model HOCs by the bacterium Sphingomonas paucimobilis EPA505 were measured at defined dissolved concentrations ranging over 4 orders of magnitude, from 0.017 to 658 µg L(-1) for phenanthrene and from 0.006 to 90.0 µg L(-1) for fluoranthene. Both compounds had similar mineralization fluxes, and these increased by 2 orders of magnitude with increasing dissolved concentrations. First-order mineralization rate constants were also similar for both PAHs, but decreased by around 2 orders of magnitude with increasing dissolved concentrations. Dynamic passive dosing is a useful tool for measuring biotransformation kinetics at realistically low and defined dissolved HOC concentrations.

A look down the drain: Identification of dissolved and particle bound organic pollutants in urban runoff waters and sediments.

Urban runoff contains a range of organic micropollutants which, if not removed during wastewater treatment, pose a risk to aquatic environments. These mixtures are complex and often site-specific. Street drains provide an ideal sampling point given they collect the runoff from local and defined catchments. In this study, runoff was collected and sampled in five street drains located in a medium sized town in Germany. A specially constructed trap was used to collect the particulate and total water fractions of the runoff. In addition, passive samplers were deployed to determine the freely dissolved concentrations of selected compounds in the runoff. In sum, 187 polar organic micropollutants could be quantified using LC-HRMS. Thirty of these could only be detected by the use of passive samplers. Traffic derived pollutants such as corrosion inhibitors, rubber- and plastic additives, but also pollutants of industrial origin were strongly represented with sum median concentrations of 100 µg/kg dry weight (DW) in the sediment and 400 ng/L in the water fraction. Several of these substances are of concern due to their environmental persistence and mobility. Perfluorinated compounds and pesticides occurred at lower levels of several µg/kg DW sediment or ng/L water. A number of substances including pharmaceuticals, sweeteners and stimulants indicated domestic wastewater influences. Furthermore, a total of 62 parent and alkylated PAHs were quantified by GC-MS and contributed 30-70% to the sum concentrations of the micropollutants. Non-EPA PAHs dominated the carcinogenic PAH toxicity. The increased PAH alkylation indices (0.7-0.9) showed these primarily came from combustion sources. The runoff particles were additionally microscopically characterized, and correlations were found between the rubber particle counts and the PAH alkylation-index as well as the levels of 2-(methylthio)benzothiazole, a marker compound for tire leaching.

Dissolved organic carbon enhances the mass transfer of hydrophobic organic compounds from nonaqueous phase liquids (NAPLs) into the aqueous phase.

The hypothesis that dissolved organic carbon (DOC) enhances the mass transfer of hydrophobic organic compounds from nonaqueous phase liquids (NAPLs) into the aqueous phase above that attributable to dissolved molecular diffusion alone was tested. In controlled experiments, mass transfer rates of five NAPL-phase PAHs (log K(OW) 4.15-5.39) into the aqueous phase containing different concentrations of DOC were measured. Mass transfer rates were increased by up to a factor of 4 in the presence of DOC, with the greatest enhancement being observed for more hydrophobic compounds and highest DOC concentrations. These increases could not be explained by dissolved molecular diffusion alone, and point to a parallel DOC-mediated diffusive pathway. The nature of the DOC-mediated diffusion pathway as a function of the DOC concentration and PAH sorption behavior to the DOC was investigated using diffusion-based models. The DOC-enhanced mass transfer of NAPL-phase hydrophobic compounds into the aqueous phase has important implications for their bioremediation as well as bioconcentration and toxicity.

Effects of the antidepressant mirtazapine on the swimming behaviour and gene expression rate of Danio rerio embryos - Is the sedating effect seen in humans also evident for fish?

In the last decade, mirtazapine has become an important antidepressant in clinical use and has also been found at many different environmental sampling sites. Several homologies between the zebrafish Danio rerio and humans, combined with a number of advantages for behavioural and gene expression research using zebrafish embryos, make their use for the analysis of mirtazapine appropriate. The sedative effect of mirtazapine in humans was also found for a specific concentration range in zebrafish embryos (1333.4 µg/L - 2666.9 µg/L). Specifically, 116 hpf old zebrafish embryos showed a reduced swimming distance when exposed to 1334.4 µg/L mirtazapine. Furthermore, changes at the gene regulatory level could be measured (1333.4 µg/L), in particular in the superordinate regulatory systems. For selected transporters of all regulatory systems, an up regulation of the genes by a factor of more than five times could be measured at the highest mirtazapine exposure concentration that was tested. Finally, studies on the protein levels demonstrated an increase in acetylcholinesterase activity for several exposure concentrations (83.3 µg/L and 666.7 µg/L). The physiological changes in zebrafish embryos caused by mirtazapine demonstrate the relevance of these types of studies in aquatic non-target organisms. Such neuroactive substances could pose a potential risk for aquatic organisms below the previously considered concentration threshold for morphological effects.

Comparing straw, compost, and biochar regarding their suitability as agricultural soil amendments to affect soil structure, nutrient leaching, microbial communities, and the fate of pesticides.

The emission of nutrients and pesticides from agricultural soils endangers natural habitats. Here, we review to which extent carbon-rich organic amendments help to retain nutrients and pesticides in agricultural soils and to reduce the contamination of surrounding areas and groundwater. We compare straw, compost, and biochar to see whether biochar outperforms the other two more traditional and cheaper materials. We present a list of criteria to evaluate the suitability of organic materials to be used as soil amendments and discuss differences in elemental compositions of straw, compost, and biochar to understand, how soil microorganisms utilize those materials. We review their effects on physical and chemical soil characteristics, soil microbial communities, as well as effects on the transformation and retention of nutrients and pesticides in detail. It becomes clear that for all three amendments their effects can vary greatly depending on numerous aspects, such as the type of soil, application rate, and production procedure of the organic material. Biochar is most effective in increasing the sorption capacity of soils but does not outperform straw and compost with regards to the other aspects investigated. Nevertheless, the possibility to design biochar properties makes it a very promising material. Finally, we provide critical comments about how to make studies about organic amendments more comparable (comprehensive provision of material properties), how to improve concepts of future work (meta-analysis, long-term field studies, use of deep-insight microbial DNA sequencing), and what needs to be further investigated (the link between structural and functional microbial parameters, the impact of biochar on pesticide efficiency).

Passive dosing for producing defined and constant exposure of hydrophobic organic compounds during in vitro toxicity tests.

Toxicity testing of hydrophobic organic compounds (HOCs) in plastic cell culture plates is problematic due to compound losses through volatilization and sorption to the wells and culture medium constituents. This leads to poorly defined exposure and reduced test sensitivity. Passive dosing can overcome these problems by the continual partitioning of HOCs from a dominating reservoir loaded in a biologically inert polymer such as silicone, providing defined and constant freely dissolved concentrations and also eliminating spiking with cosolvents. This study aimed to select a suitable passive dosing format for in vitro tests in multiwell plates and characterize its performance at 37 degrees C. Silicone O-rings were the most suitable format; they were both practical and demonstrated excellent passive dosing performance. (1) The rings were loaded by partitioning from a methanol solution containing polycyclic aromatic hydrocarbons (PAHs) (log K(OW), 3.33-6.43) that served as model compounds, followed by removal of the methanol with water. This resulted in highly reproducible HOC concentrations in the silicone O-rings. (2) The release of PAHs into aqueous solutions was rapid and reproducible, with equilibrium partitioning being reached within hours. (3) The buffering capacity of the O-rings was sufficient to maintain stable concentrations over more than 72 h. The O-rings were then applied to test a range of PAHs at their aqueous solubility in an array of established in vitro cell culture assays with human cells and cell lines. These included the formation of reactive oxygen species, induction of the IL-8 cytokine promoter, and secretion of MCP-1 by the cells. The biological responses depended on the melting point of the individual PAHs and their maximum chemical activities (a(max)). Only those PAHs with the highest a(max) stimulated the formation of reactive oxygen species and MCP-1 secretion, while they inhibited the induction of the IL-8 cytokine promoter.

Quantitative evaluation of polyethersulfone and polytetrafluoroethylene membrane sorption in a polar organic chemical integrative sampler (POCIS).

The lag effect in the polar organic chemical integrative sampler (POCIS) equipped with a polyethersulfone (PES) membrane (POCIS-PES) is a potential limitation for its application in water environments. In this study, a POCIS with a poly(tetrafluoroethylene) (PTFE) membrane (POCIS-PTFE) was investigated for circumventing membrane sorption in order to provide more reliable concentration measurements of organic contaminants. Sampler characteristics such as sampling rates (RS) and sampler-water partition coefficients (KSW) were similar for POCIS-PES and POCIS-PTFE, indicating that partitioning into Oasis HLB as the receiving phase dominates the overall partitioning from the aqueous phase to the POCIS. Membrane sorption was quantified in both laboratory and field experiments. Although POCIS-PTFE showed minor membrane sorption, the PTFE membranes were not robust enough to prevent changes in the sorption of the pollutants to the inner Oasis HLB sorbent due to biofouling. This was reflected in significant ionization effects in the electrospray ionization (ESI) source during the LC-MS/MS analysis. Despite clear differences in the ionization effects, the two POCISs types provided similar time-weighted average (CTWA) concentrations after a two-week passive sampling campaign in surface water and the outflow of a wastewater treatment plant. This study contributes to a more detailed understanding of POCIS application by providing a quantitative evaluation of membrane sorption and its associated effects in the laboratory and field.

Do you smell the danger? Effects of three commonly used pesticides on the olfactory-mediated antipredator response of zebrafish (Danio rerio).

Fish are warned about the presence of predators via an alarm cue released from the skin of injured conspecifics. The detection of this odor inherently initiates an antipredator response, which increases the chance of survival for the individual. In the present study, we assessed the effect of three commonly used pesticides on the antipredator response of zebrafish (Danio rerio). For this, we analyzed the behavioral response of zebrafish to a conspecific skin extract following 24 h of exposure to the respective contaminants. Results demonstrate that fish exposed to 20 µg/L of the organophosphate insecticide chlorpyrifos significantly reduced bottom-dwelling and freezing behavior, suggesting an impairment of the antipredator response. For the urea-herbicide linuron and the pyrethroid insecticide permethrin, no statistically significant effects could be detected. However, linuron-exposed fish appeared to respond in an altered manner to the skin extract; some individuals failed to perform the inherent behaviors such as erratic movements and instead merely increased their velocity. Furthermore, we determined whether zebrafish would avoid the pesticides in a choice maze. While fish avoided permethrin, they behaved indifferently to chlorpyrifos and linuron. The study demonstrates that pesticides may alter the olfactory-mediated antipredator response of zebrafish in distinct ways, revealing that particularly fish exposed to chlorpyrifos may be more prone to predation.