[Show abstract][Hide abstract] ABSTRACT: ECHA’s topical scientific workshops facilitate dialogue between academia, regulators, the regulated industry and other stakeholders in order to assess the regulatory impact of the latest scientific developments in a particular area. The aim is that new or improved approaches of practical use in REACH, CLP and the Biocidal
Products Regulation will emerge, catalysed by the discussions and emerging common understandings in the workshops.
Following a proposal from a group of experts, ECHA organised the first Topical Scientific Workshop on Risk Assessment for the Sediment Compartment.
The Topical Scientific Workshop on Risk Assessment for the Sediment Compartment was held from 7 to 8 May 2013 at ECHA, bringing together experts in the field of sediment risk assessment to brainstorm and develop updated scientific principles and guidelines for assessing ecological risks of chemical substances for freshwater and marine sediments. Recent developments for particular substance types (such as metals)
and a broad understanding of risk assessment methodologies in other regulated products and from schemes outside the EU were part of the discussion.
The primary objective of this workshop was to review the state of the art and to develop updated scientific principles and guidelines for assessing ecological risks of chemical substances for freshwater and marine sediments. The discussion elements included:
• Discussing the current state of the science on sediment toxicology.
• Reviewing current risk assessment frameworks for sediments relying on the extrapolation from the pelagic community, and developing further recommendations on the applicability of these extrapolation approaches.
• Addressing the water-sediment interface and the epi-benthonic community.
• Establishing links among available lines of laboratory and field evidence on ecotoxicity, bioavailability, and ecosystem quality/function, and specifically for freshwater systems.
• Developing general principles applicable to different regulatory schemes, considering the protection goals set forth by current regulatory processes whilst focusing on the regulation of chemicals underREACH and CLP. The broader context is biocides, plant protection products and pharmaceuticals, and broader legal instruments are also relevant, e.g. the Water Framework Directive, the Industrial Emissions Directive, and equivalent regulatory processes in non-EU jurisdictions. The workshop brought together over 100 experts from around the world to set the scientific principles for assessing risks to the sediment compartment in all regulatory contexts. The two-day workshop included
general plenary sessions with case studies and topical breakout group sessions, where the participants discussed specific recommendations on how to use scientific knowledge for regulatory purposes.
[Show abstract][Hide abstract] ABSTRACT: REACH(1) entered into force in June 2007 and has hence been operational for six years. With the first registration deadline in November 2010, the European Chemicals Agency (ECHA(2)) has received a large amount of scientific and administrative information related to chemical substances. In order to understand what type of data on ecotoxicity endpoints was submitted under the REACH framework a detailed analysis of the availability and content of relevant information was performed. To avoid unnecessary testing, the REACH Regulation provides registrants with the possibility to build testing strategies and to adopt the standard information requirements based on the specific conditions listed in the regulation. The types of information submitted by registrants to fulfil data requirements for aquatic, sediment and terrestrial toxicity endpoints were analysed. The REACH database analysis confirms large differences in the availability of experimental aquatic versus sediment and soil ecotoxicity data. Information requirements on aquatic organisms are mainly covered by experimental data, while those for sediment and soil are mostly waived.
Science of The Total Environment 11/2013; 470-471C:1225-1232. · 3.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This paper summarises the terrestrial ecotoxicity data submitted in the REACH(1) registration dossiers and disseminated by ECHA.(2) The analysis describes both the guidelines and the test species mostly used by registrants. REACH information requirements in relation to the effects on terrestrial organisms encompass three trophic levels; invertebrates, plants and micro-organisms, and the study of both long and short-term exposure. The results observed for soil invertebrates showed that on one hand there was a clear prevalence for testing on the species recommended by the standard test guidelines. On the other, the reporting included a large variety of species from very different families, demonstrating the feasibility for conducting toxicity tests on a number of relevant groups e.g. for species sensitivity distribution approaches. Standard toxicity testing with terrestrial plants under REACH follows a different approach and requires simultaneous testing on several species, using the same test conditions, adapted to each species, if needed. The test methods used to conduct the studies were only reported for 30% of cases. The most extensively reported test guidelines for terrestrial plants were OECD(3) 208, ISO(4) 11269-1 and ISO 11269-1. Information requirements for soil micro-organisms under REACH are related to the analysis of functional endpoints instead of on species or taxa. As recommended in REACH, OECD 216 and OECD 217 were the most often used test methods for soil micro-organisms. But overall, the test method was reported for only about 40% of the experimental studies. Moreover, it is noted that information on potential effects on soil micro-organisms is available for a limited number of REACH registered substances. The assessment suggests that providing waiving justifications and collecting available information, which in many cases might be well used for covering standard REACH data requirements, have been the main approaches used by registrants for the first REACH registration deadline.
Science of The Total Environment 11/2013; · 3.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Standardized test protocols are used in the regulatory context for identifying the hazardous properties of chemicals, wastes, and contaminated materials. This paper compares the relevance of two guidelines measuring effects on terrestrial plants, the OECD TG 208 and the ISO TG 22030 and presents the scientific basis for a recent decision of the European Chemicals Agency (ECHA) under the European chemicals regulation REACH. If there are no specific phytotoxicity alerts, both guidelines are considered suitable for assessing long-term hazards, providing that a sufficient number of species is included in the OECD protocol, the recommended minimum number is six, which offer a reasonably broad selection of species to account for interspecies sensitivity. The proposed methodology, based on a combination of probabilistic assessments using Monte Carlo analysis, can be adapted for supporting similar decisions under specific regulatory processes; for example, for assessing contaminated soils or pesticides' applications.
[Show abstract][Hide abstract] ABSTRACT: The exposure to benzo[a]pyrene (B[a]P) results in an alteration of immune function in mammals and fish, and the analysis of cytokine mRNA levels has been suggested for predicting the immunomodulatory potential of chemicals. To obtain evidence of the innate immune responses to B[a]P in Xenopus laevis, the present study monitored the mRNA expression of interleukin 1-β (IL-1β), tumor necrosis factor α (TNF-α) and heat shock protein 70 (HSP70) in a laboratorial exposure. Tadpoles exposed to 8.36, 14.64, 89.06 and 309.47 μg/L of B[a]P,were used for detecting hsp70, IL-1β and TNF-α mRNA induction. A dose-response increase in the expression of hsp70 and IL-1β mRNA was found. The results of this study confirmed the use of hsp70 and IL-1β, but not TNF-α, as sensitive indicators of immunotoxic effect of B[a]P in X. laevis. Further research would be required for the validation of these endpoints.
[Show abstract][Hide abstract] ABSTRACT: Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC(50)) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians.
The Scientific World Journal 01/2012; 2012:605804. · 1.73 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Phytoremediation and bioremediation are site-specific processes, and feasibility studies should be performed as an initial step in scaling-up these processes. Soil microcosms can be a useful tool for investigating the efficiency of remediation alternatives under realistic conditions. In the present study, the degradation of diesel fuel in soil was studied in artificially assembled microcosms during a 180-day experiment. Microcosms consisted of soil columns seeded with a mix of Festuca arundinacea and Trifolium pratense into which earthworms were introduced and native soil microflora was used. Remediation assays were performed with different combinations of soil organisms' assemblages (micro-organisms, earthworms and plants) to determine the effects of these organisms and their interaction on diesel degradation. The microcosm system allowed for the study of the efficiency of remediation under field-relevant conditions. This system provided information about the dissipation of the pollutants as well as chemical leaching and possible toxic effects on the organisms during the remediation process. The decrease in soil hydrocarbon levels depended on the organisms' assemblage. Plants were not effective in the remediation of these soils despite their stimulation of microbial biomass. Conversely, earthworms had a beneficial impact on the dissipation of hydrocarbons that did not appear to be related to a generic improvement of plant or microbial activity. The grass species was more tolerant than the legume species to diesel fuel-contaminated soils. Leaching of hydrocarbons was negligible and independent of the organisms' assemblage.
Ecotoxicology and Environmental Safety 09/2011; 74(8):2133-40. · 2.20 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Leachates from municipal solid waste (MSW) landfills may contain a huge diversity of contaminants; these wastewaters should be considered as potentially hazardous complex mixtures, representing a potential environmental risk for surface and groundwater. Current MSW landfill wastes regulatory approaches deem exclusively on the physicochemical characterization and does not contemplate the ecotoxicological assessment of landfill leachates. However, the presence of highly toxic substances in consumer products requires reconsideration on the need of more specific ecotoxicological assessments. The main aim of this study was to evaluate the toxicity of different MSW landfill leachates using a battery of toxicity tests including acute toxicity tests with Daphnia magna and the anuran Xenopus laevis and the in vitro toxicity test with the fish cell line RTG-2. The additional objective was to study the possible correlation between physicochemical properties and the toxicity results obtained for untreated landfill leachates. The results showed that the proposed test battery was effective for the ecotoxicological characterization of MSW landfill leachates. A moderate to strong correlation between the measured physicochemical parameters and the calculated toxicity units was detected for all toxicity assays. Correlation factors of 0.85, 0.86 and 0.55 for Daphnia, Xenopus and RTG-2 tests, respectively, were found. The discriminant analysis showed that certain physicochemical parameters could be used for an initial categorization of the potential aquatic acute toxicity of leachates; this finding may facilitate leachates management as the physicochemical characterization is currently the most common or even only monitoring method employed in a large majority of landfills. Ammonia, alkalinity and chemical oxygen demand (COD), together with chloride, allowed a proper categorization of leachates toxicity for up to 75% of tested samples, with a small percentage of false negatives.
[Show abstract][Hide abstract] ABSTRACT: This study aimed to characterize quantitatively the temporal basal and induced ethoxyresorufin-O-deethylase (EROD) activity as indicator of cytochrome P4501A (CYP1A) function during embryonic development of medaka (Oryzias latipes). For this purpose, non-invasive methods over fluorescence images of the whole embryo (non-organ-specific [NOS] EROD activity) or specifically of the gallbladder (organ-specific [OS] EROD activity) were used. To induce this EROD activity, embryos were continuously exposed to β-naphthoflavone (BNF; 0.005, 0.05, 0.5, 5 μg/L). Analytical chemistry suggested no signs of BNF dissipation. Mean fluorescence intensity values for EROD induction increased with BNF concentration throughout embryonic development. Significant increments in the NOS activity were seen from exposures to ≥ 0.5 μg BNF/L as early as 2 days post-fertilization (dpf), and in the OS EROD activity as soon as the gallbladder was conspicuous (i.e. 4 dpf). Morphometric in vivo analysis of the gallbladder during embryonic development did not indicate significant dilation after BNF treatment suggesting normal hepatic bile formation. The conditions optimized in this study using intact embryos should allow the quantitation of EROD activity induced by specific chemicals, mixtures and environmental samples in terms of BNF-equivalents, offering a proper estimation of their potency. These results demonstrate the utility of medaka in a fish embryo test for a non-invasive CYP1A analysis expressed as EROD activity, fitting in the three R principles for the minimization of animal use in ecotoxicology evaluations and that are among the objectives of the European Community regulation for the Registration, Evaluation, Authorization and Restriction of Chemical substances (REACH).
[Show abstract][Hide abstract] ABSTRACT: The antiparasitic ivermectin is of particular concern to regulatory agencies. Ivermectin can reach the environment through the direct emission of dung from livestock on pasture and via manure application on agricultural lands.
A semifield study was conducted for assessing the ivermectin dynamic in runoff and drainage waters from dung-treated soils placed on experimental trays. The experiment was conducted under natural Mediterranean conditions. Realistic pasture and arable land applications were assessed using dung of treated animals and compared with a positive control (spraying the ivermective solution without dung).
Similar concentrations were obtained in all three treatments for drainage waters, with values ranging from <5-10 to about 20 ng/l. However, strong treatment-related variation was observed in runoff waters, with the highest concentrations found in the spray treatment (9-188 ng/l), followed by the arable land (<5-88 ng/l) scenario, and concentrations not exceeding 6 ng/l in the pasture scenario. Ivermectin levels in runoff particles were up to 1,660 and 5,890 ng/kg dry weight for the pasture (I1) and arable land (I2) scenarios, respectively. Ivermectin was only detected in the drainage and runoff waters collected in the first rainfall events after treatment.
The measured concentrations in water (0.006-0.118 ng/ml) and runoff particles (0.052-5.89 ng/mg dry suspended matter) are orders of magnitude higher than those provoking effects on aquatic and benthonic communities under experimental and mesocosm conditions, suggesting a clear risk for aquatic systems in the vicinity of pasture areas of treated animals or arable soil fertilized with its manure.
Environmental Science and Pollution Research 03/2011; 18(7):1194-201. · 2.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The environmental risk assessment of the veterinary pharmaceutical ivermectin is receiving significant attention. This paper assesses the capacity of the MS·3 soil microcosm as a tool for targeting the environmental impact assessment of veterinary drugs, using ivermectin as model. Two screening MS·3 were performed using different European soils; one with a soil collected in an agricultural station near to Madrid, Spain and a second with a soil collected in a farm area close to York, UK. Soils were fortified with ivermectin at the following ranges: 0.01-10 mg kg–1 and 0.1-100 mg kg–1 in the Madrid and York studies, respectively. The effects on earthworms, plants and soil microorganisms were assessed in the Madrid soil. Toxicity tests on aquatic organisms (algae, cladocerans and in vitro fish cell line RTLW1) were also conducted with the leachates. No effects were observed in earthworms and plants at any tested concentration; reduction in the respiration rate (< 5%) of soil microorganisms was detected. Earthworm/soil bioconcentration factors decreased with the increase in soil concentrations and were higher for the York soil. Effects on daphnids were observed in tested leachates; based on measured levels of ivermectin in the leachates an EC50 of about 0.5 μgL–1 can be estimated. Comparisons based on toxicity data and equilibrium partitioning confirmed that the main risk is expected to be related to the high sensitivity of cladocerans. The results confirm that MS·3 systems are cost-effective tools for assessing the impact of veterinary pharmaceuticals when applied to agricultural land, as previously demonstrated for antimicrobials.
SPANISH JOURNAL OF AGRICULTURAL RESEARCH 01/2011; 9(2):433-443. · 0.66 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This paper describes a new methodology for assessing site-specific environmental impact of contaminants. The proposed method integrates traditional risk assessment approaches with real and variable environmental characteristics at a local scale. Environmental impact on selected receptors was classified for each environmental compartment into 5 categories derived from the whole (chronic and acute) risk assessment using 8 risk levels. Risk levels were established according to three hazard quotients (HQs) which represented the ratio of exposure to acute and chronic toxicity values. This tool allowed integrating in only one impact category all the elements involved in the standard risk assessment. The methodology was applied to an abandoned metal mine in Spain, where high levels of As, Cd, Zn and Cu were detected. Risk affecting potential receptors such as aquatic and soil organisms and terrestrial vertebrates were assessed. Whole results showed that impact to the ecosystem is likely high and further investigation or remedial actions are necessary. Some proposals to refine the risk assessment for a more realistic diagnostic are included.
Science of The Total Environment 01/2011; 409(4):692-703. · 3.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: High-resolution gas chromatography/mass spectrometry (HRGC/MS) is the standard method for analysing dioxin, furan and polybrominated retardants in hazardous waste. Determination of dioxin-like compounds using in vitro bioassays such as ethoxyresorufin-O-deethylase (EROD) is an important tool to evaluate their Ah receptor-mediated toxic effects, because it detects all arylhydrocarbon receptor ligands in a variety of sample matrices. In the present work, we compared RTG-2 cell line EROD bioassay with HRGC/MS for assessing waste samples (liquid and solid) contaminated with polychlorinated dibenzo-p-dioxins and dibenzofurans, polychlorinated biphenyls (dioxin-like PCBs) and other xenobiotics. For liquid samples, HRGC/MS-toxic equivalent (HRGC/MS-TEQ) values ranged from 273.26 to 5.84 ng TEQ l(-1) and correlated well (correlation coefficient 0.99) with values obtained by EROD-TEQ, which ranged from 128 to 2.5 ng TEQ l(-1). For solid samples, HRGC/MS-TEQ values ranged from 3.44 to 0.49 ng TEQ g(-1) and correlated less well than liquid samples (correlation coefficient 0.64) with values obtained by EROD-TEQ ranging from 2.27 to 0.93 ng TEQ g(-1). The overestimation of RTG-2 EROD-TEQ (1.2 +/- 0.92 of values established by HRGC/MS) and the absence of false-negative results may limit analytical costs by eliminating the need for follow-up GC/MS analysis on the negative samples. We suggest that RTG-2 EROD bioassay is an inexpensive means for preliminary dioxin and furan positive screenings of waste samples.
Journal of Applied Toxicology 08/2010; 30(6):603-10. · 2.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Screening whole effluent toxicity tests are cost/effective methods for detecting the presence of toxic concentrations of unknown pollutants, but the application must solve the problem associated with the effect of high and variable concentrations of nutrients in the effluent on the results of algal toxicity tests. This work proposes a cost/effective test, based on three dilution levels measured at a single point time and a discriminant model for establishing if this kind of complex samples, with difficult interpretation of dilution-response curves, should be considered toxic to algae. This procedure identified properly around 85% of the samples considered toxic by expert judgement.
Bulletin of Environmental Contamination and Toxicology 07/2010; 85(1):72-8. · 1.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The serotonin re-uptake inhibitor fluoxetine was selected for an environmental risk assessment, using the most recent European guideline (EMEA 2006) within the European Union (EU)-funded Environmental Risk Assessment of Pharmaceuticals (ERAPharm) project due to its environmental persistence, acute toxicity to nontarget organisms, and unique pharmacokinetics associated with a readily ionizable compound. As a widely prescribed psychotropic drug, fluoxetine is frequently detected in surface waters adjacent to urban areas because municipal wastewater effluents are the primary route of entry to aquatic environments. In Phase I of the assessment, the initial predicted environmental concentration of fluoxetine in surface water (initial PEC(SW)) reached or exceeded the action limit of 10 ng/L, when using both a default market penetration factor and prescription data for Sweden, Germany, and the United Kingdom. Consequently, a Phase II risk assessment was conducted in which green algae were identified as the most sensitive species with a NOEC of <0.6 microg/L. From this value, a predicted no effect concentration for surface waters (PNEC(SW)) of 0.012 microg/L was derived. The PEC/PNEC ratio was above the trigger value of 1 in worst-case exposure scenarios indicating a potential risk to the aquatic compartment. Similarly, risks of fluoxetine for sediment-dwelling organisms could not be excluded. No risk assessment was conducted for the terrestrial compartment due to a lack of data on effects of fluoxetine on soil organisms. The need for a separate risk assessment for the main metabolite of fluoxetine, norfluoxetine, was not conducted because of a lack of fate and effect studies. Based on published data, fluoxetine and norfluoxetine appeared to have a low to moderate bioaccumulation potential, which should be confirmed in formal studies according to OECD guidelines. Exposure assessments for fluoxetine according to the current framework rely heavily on K(OC) and K(OW) values. This approach is problematic, because fluoxetine is predominantly a cationic substance at environmental pH values. Consequently, the fate of fluoxetine (and other ionic substances) cannot be predicted using partition coefficients established for nonionic compounds. Further, published estimates for partition coefficients of fluoxetine vary, resulting in considerable uncertainties in both the exposure and environmental risk assessments of fluoxetine.
Integrated Environmental Assessment and Management 07/2010; 6 Suppl:524-39.
[Show abstract][Hide abstract] ABSTRACT: The veterinary parasiticide ivermectin was selected as a case study compound within the project ERAPharm (Environmental Risk Assessment of Pharmaceuticals). Based on experimental data generated within ERAPharm and additional literature data, an environmental risk assessment (ERA) was performed mainly according to international and European guidelines. For the environmental compartments surface water, sediment, and dung, a risk was indicated at all levels of the tiered assessment approach. Only for soil was no risk indicated after the lower tier assessment. However, the use of effects data from additional 2-species and multispecies studies resulted in a risk indication for collembolans. Although previously performed ERAs for ivermectin revealed no concern for the aquatic compartment, and transient effects on dung-insect populations were not considered as relevant, the present ERA clearly demonstrates unacceptable risks for all investigated environmental compartments and hence suggests the necessity of reassessing ivermectin-containing products. Based on this case study, several gaps in the existing guidelines for ERA of pharmaceuticals were shown and improvements have been suggested. The action limit at the start of the ERA, for example, is not protective for substances such as ivermectin when used on intensively reared animals. Furthermore, initial predicted environmental concentrations (PECs) of ivermectin in soil were estimated to be lower than refined PECs, indicating that the currently used tiered approach for exposure assessment is not appropriate for substances with potential for accumulation in soil. In addition, guidance is lacking for the assessment of effects at higher tiers of the ERA, e.g., for field studies or a tiered effects assessment in the dung compartment.
Integrated Environmental Assessment and Management 07/2010; 6 Suppl:567-87.
[Show abstract][Hide abstract] ABSTRACT: The current study describes the effect of cypermethrin, fluoxetine, and thiabendazole, at environmentally relevant concentrations, on the expression of heat shock protein 70 (HSP70) and interleukin 1β (IL-1β), using Xenopus laevis larvae as animal model. Cytokines and interleukins are considered good predictors of the immunotoxic potential of xenobiotics. Tadpoles at stage 47 (normal tables of X. laevis) were exposed under static conditions to: 0.3 and 30 µg/L fluoxetine, 0.7 µg/L thiabendazole, and 0.24 µg/L cypermethrin. The effects were evaluated at 7, 24, and 72 h, and 6 and 9 d. Randomly chosen tadpoles were used as genetic material for detection of hsp70 and IL-1β mRNA induction through reverse transcription PCR. Tadpoles exposed to 30 µg/L fluoxetine showed mRNA expression of both genes at all exposure times, whereas at 0.3 µg/L a peak response for hsp70 was observed after 24 h, and the increase in IL-1β mRNA was statistically significant with respect to the control 72 h after exposure. Thiabendazole induced a high expression of mRNA for both hsp70 and IL-1β at all exposure times. Cypermethrin increased the hsp70 mRNA levels, with a peak at 24 h, and provoked high expression of IL-1β mRNA at all exposure times. Considering the relationship between HSP70 and IL-1β and their involvement (mainly of IL-1β) in immune responses, certain changes observed in their expression could be considered warning indicators of potential immunotoxic effects of these substances on Xenopus.
Environmental Toxicology and Chemistry 07/2010; 29(11):2536-43. · 2.62 Impact Factor