Data ranges in aquatic toxicity of chemicals: Consequences for environmental risk analysis

A significant problem for effect assessment of aquatic ecosystems arises from the large ranges of toxicity data, which can be found in different databases and literature. Here, ranges are given for the aquatic toxicity of 27 high production volume chemicals. Based on these illustrative examples and on the current literature on uncertainty in aquatic effect assessment, toxicity ranges are discussed for their possible causes (variation in experimental condition, species, endpoint, time) and ways to handle them (safety factors). Implications and recommendations on the procedure of risk analysis of chemical substances are drawn. Two main requirements for a comprehensive risk assessment are identified, which often play a minor role in current practice (as they are often neglected) as well as in scientific discussion (as they are meant to be trivial). First, data quality must be checked critically before applying any result of a toxicity test. Secondly, experimental data should take into account different species and acute as well as chronic data. If these aspects are considered in risk analysis, which is common practice in ecotoxicology but not always in the context of practical applications in risk engineering, a more comprehensive picture of the environmental toxicity of a chemical substance can be obtaine

The Ultraviolet Filter 3-Benzylidene Camphor Adversely Affects Reproduction in Fathead Minnow (Pimephales promelas)

The ultraviolet (UV) filter 3-benzylidene camphor (3BC) is used in personal care products and in a number of materials for UV protection. 3BC has been shown in vitro and in vivo in fish to be estrogenic, but possible effects on fertility and reproduction are unknown. In this study we evaluate whether 3BC affects reproduction of fish Pimephales promelas. After a preexposure period of 21 days, reproductively mature fathead minnows were exposed to increasing concentrations of 3BC for 21 days in a static-renewal procedure. Actual 3BC concentrations decreased to 23% of initial levels and median concentrations were 0.5, 3, 33, 74, and 285 μg/l. 3BC affected reproduction in a dose-dependent manner with weak effects on fecundity at 3 μg/l, a significant decrease at 74 μg/l, and a cessation of reproduction at 285 μg/l. 3BC was accumulated in fish with an average bioconcentration factor of 313 ± 151. Dose-dependent demasculinization in secondary sex characteristics of male fish and dose-dependent induction of plasma vitellogenin occurred, which was significant at 74 μg/l and higher. 3BC had a profound and dose-dependent effect on the histology of gonads of male and female fish at 3 μg/l and higher. At 74 and 285 μg/l, oocyte and spermatocyte development was inhibited in male and female gonads. Testes of exposed males had much fewer spermatogenic cysts, and ovaries of exposed females had much fewer mature but more atretic, follicles. This study shows significant effects of the UV filter 3BC on fertility, gonadal development, and reproduction of fish after short-term exposure that may have negative consequences on the population leve

Comparison of In Vitro and In Vivo Estrogenic Activity of UV Filters in Fish

In this work, we evaluate whether in vitro systems are good predictors for in vivo estrogenic activity in fish. We focus on UV filters being used in sunscreens and in UV stabilization of materials. First, we determined the estrogenic activity of 23 UV filters and one UV filter metabolite employing a recombinant yeast carrying the estrogen receptor of rainbow trout (rtERα) and made comparisons with yeast carrying the human hERα for receptor specificity. Benzophenone-1 (BP1), benzophenone-2 (BP2), 4,4-dihydroxybenzophenone, 4-hydroxybenzophenone, 2,4,4-trihydroxy-benzophenone, and phenylsalicylate showed full dose-response curves with maximal responses of 81-115%, whereas 3-benzylidene camphor (3BC), octylsalicylate, benzylsalicylate, benzophenone-3, and benzophenone-4 displayed lower maximal responses of 15-74%. Whereas the activity of 17β-estradiol was lower in the rtERα than the hERα assay, the activities of UV filters were similar or relatively higher in rtERα, indicating different relative binding activities of both ER. Subsequently, we analyzed whether the in vitro estrogenicity of eight UV filters is also displayed in vivo in fathead minnows by the induction potential of vitellogenin after 14 days of aqueous exposure. Of the three active compounds in vivo, 3BC induced vitellogenin at lower concentrations (435 μg/l) than BP1 (4919 μg/l) and BP2 (8783 μg/l). The study shows, for the first time, estrogenic activities of UV filters in fish both in vitro and in vivo. Thus we propose that receptor-based assays should be used for in vitro screening prior to in vivo testing, leading to environmental risk assessments based on combined, complementary, and appropriate species-related assays for hormonal activit

Identification of a CYP3A form (CYP3A126) in fathead minnow ( Pimephales promelas ) and characterisation of putative CYP3A enzyme activity

Cytochrome P450-dependent monooxygenases (CYPs) are involved in the metabolic defence against xenobiotics. Human CYP3A enzymes metabolise about 50% of all pharmaceuticals in use today. Induction of CYPs and associated xenobiotic metabolism occurs also in fish and may serve as a useful tool for biomonitoring of environmental contamination. In this study we report on the cloning of a CYP3A family gene from fathead minnows (Pimephales promelas), which has been designated as CYP3A126 by the P450 nomenclature committee (GenBank no. EU332792). The cDNA was isolated, identified and characterised by extended inverse polymerase chain reaction (PCR), an alternative to the commonly used method of rapid amplification of cDNA ends. In a fathead minnow cell line we identified a full-length cDNA sequence (1,863 base pairs (bp)) consisting of a 1,536bp open reading frame encoding a 512 amino acid protein. Genomic analysis of the identified CYP3A isoenzyme revealed a DNA sequence consisting of 13 exons and 12 introns. CYP3A126 is also expressed in fathead minnow liver as demonstrated by reverse transcription PCR. Exposure of fathead minnow (FHM) cells with the CYP3A inducer rifampicin leads to dose-dependent increase in putative CYP3A enzyme activity. In contrast, inhibitory effects of diazepam treatment were observed on putative CYP3A enzyme activity and additionally on CYP3A126 mRNA expression. This indicates that CYP3A is active in FHM cells and that CYP3A126 is at least in part responsible for this CYP3A activity. Further investigations will show whether CYP3A126 is involved in the metabolism of environmental chemicals. Figure Induction of CYP3A activity by rifampicin and inhibition by diazepam in FHM cell

Molecular Mechanisms in Ecotoxicology: An Interplay between Environmental Chemistry and Biology

A close collaboration between environmental chemistry and biological sciences is required for a complete understanding of ecotoxicological effects. Bioavailability and uptake of pollutants cannot be regarded as isolated chemical or biological questions. Knowledge of the effective concentrations in the organism or at the target site(s) is essential to link the fate and effects of a chemical and is a prerequisite for quantitative investigation of the modes of toxic action. These modes of action need to be unraveled using whole-organism or in vitro systems in order to be able to develop specific biomarkers and biosensors that can be applied as early warning systems. Our mode-of-action-based approaches, in which chemical and biological analytical tools are combined, should improve the understanding of ecotoxicological effects and should be implemented in the future in risk assessment

Effects of the Protein Kinase Inhibitor PKC412 on Gene Expression and Link to Physiological Effects in Zebrafish Danio rerio Eleuthero-Embryos

To identify molecular effects of the antineoplastic agent protein kinase C inhibitor 412 (PKC412) (midostaurin), we applied gene expression profiling in zebrafish using whole-genome microarrays. Behavioral, developmental, and physiological effects were investigated in order to analyze for correlations between altered gene expression profiles with effects on development and physiology. Zebrafish blastula-stage embryos were exposed for 6 days postfertilization to nominal levels of 2 and 40 μg/l PKC412. Among the 259 and 511 altered transcripts at both concentrations, respectively, the expressions of genes involved in the circadian rhythm were further investigated. Alteration of swimming behavior was not observed. Pathways of interest affected by PKC412 were angiogenesis, apoptosis, DNA damage response, and response to oxidative stress. Angiogenesis was analyzed in double-transgenic zebrafish embryos Tg(fli1a:EGFP)y1;Tg(gata1:dsRed)sd2; no major defects were induced by PKC412 treatment at both concentrations. Apoptosis occurred in olfactory placodes of embryos exposed to 40 μg/l, and DNA damage was induced at both PKC412 concentrations. However, there were no significant effects on reactive oxygen species formation. This study leads to the conclusion that PKC412-induced alterations of gene transcripts are partly paralleled by physiological effects at high, but not at low PKC412 concentrations expected to be of environmental relevanc

The Fate of Organic Pollutants in Wastewater and Sludge Treatment: Significant Processes and Impact of Compound Properties

The fate of organic pollutants during wastewater and sludge treatment is determined by three main processes: gas exchange, sorption to suspended solids, and biodegradation. The influence of these processes differs strongly depending on the physicochemical properties of the individual compound and the particular treatment stage. For the assessment of the fate of trace pollutants in wastewater treatment, the impact of these processes must be evaluated. An acceptable removal in mechanical-biological wastewater treatment is achieved for hydrophilic compounds if they are rapidly degradable under aerobic conditions. Substances with lipophilic or amphiphilic properties should be degradable under aerobic and anaerobic conditions in order to prevent accumulation in digested sewage sludges. This article presents recent and current investigations at EAWAG which deal with the fate of selected organic substances in municipal wastewater and sludge treatment