Review of the environmental fate and effects of two UV filter substances used in cosmetic products.

Sunscreens containing UV filters, such as octocrylene (OCR) and butyl-methoxydibenzoylmethane (BMDBM), have been increasingly used to protect human skin against UV radiation. Both substances have been detected in monitoring studies in the freshwater and marine environment, and there has been concern about potential effects on aquatic organisms. In the present work, the environmental fate and occurrence, bioaccumulation and ecotoxicity including endocrine effects of OCR and BMDBM are reviewed focusing on the aquatic environment. The two UV filters have low water solubilities and a high sorption potential. The available data indicate that OCR is poorly biodegradable. BMDBM lacks anaerobic and inherent biodegradability. However, it was biodegraded to variable degrees in simulation studies. Measured concentrations in the freshwater and marine environment were found to vary considerably between sites, depending on the extent of recreational activities or wastewater discharges. While the bioconcentration factor of OCR in fish is below the threshold value for bioaccumulation according to EU REACH, the available data for BMDBM do not allow a definitive conclusion on its bioaccumulation potential. Analysis of the aquatic toxicity data showed that data quality was often limited, e.g. in the case of effect concentrations substantially exceeding maximum achievable dissolved concentrations. Up to their limit of water solubility, OCR and BMDBM showed no toxicity to microorganisms, algae, and corals, and no acute toxicity to daphnids and fish. In chronic daphnid tests, OCR was highly toxic, whereas BMDBM lacked toxicity. Reliable water-sediment toxicity tests are required to further evaluate possible effects on benthic invertebrates. The available data do not provide evidence for endocrine effects of the two UV filters on fish. In order to assess potential environmental risks caused by OCR and BMDBM, a validated exposure model for estimating direct emission of UV filters into the aquatic environment and data from systematic, longer-term monitoring studies are needed.

Environmental risk or benefit? Comprehensive risk assessment of groundwater treated with nano Fe0-based Carbo-Iron®.

Groundwater is essential for the provision of drinking water in many areas around the world. The performance of the groundwater-bearing aquifer relies on the ecosystem services provided by groundwater-related organisms. Therefore, if remediation of contaminated groundwater is necessary, the remediation method has to be carefully selected to avoid risk-risk trade-offs that might impact these ecosystems. In the present study, the environmental risk of the in situ remediation agent Carbo-Iron was performed. Carbo-Iron® is a composite of zero valent nano-iron and active carbon. Existing ecotoxicity data were complemented by studies with Daphnia magna (Crustacea), Scenedesmus vacuolatus (Algae), Chironomus riparius (Insecta) and nitrifying soil microorganisms. The predicted no effect concentration of 0.1 mg/L was derived from acute and chronic ecotoxicity studies. It was compared to measured and modelled environmental concentrations of Carbo-Iron applied in a groundwater contaminated with chlorohydrocarbons in a field study and risk ratios were derived. A comprehensive assessment approach was developed further based on existing strategies and used to identify changes of the environmental risk due to the remediation of the contaminated site with Carbo-Iron. With the data used in the present study, the total environmental risk decreased by approximately 50% in the heavily contaminated zones after the application of Carbo-Iron. Thus, based on the results of the present study, the benefit of remediation with Carbo-Iron seems to outweigh its negative effects on the environment.

Proposal for a Monitoring Concept for Veterinary Medicinal Products with PBT Properties, Using Parasiticides as a Case Study.

The aim of this work is to prepare a proposal for the post-authorization monitoring (PAM) of veterinary medicinal products (VMP), in particular parasiticides. Such a monitoring might especially be useful for parasiticides identified as Persistence Bioaccumulation Toxicity (PBT) substances, i.e., chemicals that are toxic (T), persist in the environment (P) and bioaccumulate (B) in food chains and, thus, pose a hazard to ecosystems. Based on a literature search, issues to be considered when performing such a PAM are discussed (e.g., residue analysis, compartments to be included, selection of organisms and the duration of monitoring studies). The outcome of this discussion is that-and despite that there are huge challenges in detail (e.g., in terms of analytical chemistry or taxonomy)-the technical performance of such a PAM is not the main problem, since most of the chemical and biological methods to be used are well-known (partly even standardized) or could be adapted. However, it is very difficult to define in detail where and when a monitoring should be performed. The main problem is to link exposure to effects of a certain parasiticide in a way that any impact can directly be related to the use of this parasiticide. Therefore, a "Targeted Environmental Monitoring" (TEM) is proposed, which is essentially a combination between a field study and a PAM.

Oxidized Carbo-Iron causes reduced reproduction and lower tolerance of juveniles in the amphipod Hyalella azteca.

For in situ remediation of groundwater contaminated by halogenated hydrocarbons Carbo-Iron®, a composite of microscale activated carbon and nano Fe0, was developed. Against the background of intended release of Carbo-Iron into the environment in concentrations in the g/L-range, potential ecotoxicological consequences were evaluated in the present study. The nano Fei0 in Carbo-Iron acts as reducing agent and is oxidized in aqueous systems by chlorinated solvents, groundwater constituents (e.g. dissolved oxygen) and anaerobic corrosion. As Carbo-Iron is generally oxidized rapidly after application into the environment, the oxidized state is environmentally most relevant, and Carbo-Iron was used in its oxidized form in the ecotoxicological tests. The amphipod Hyalella azteca was selected as a surrogate test species for functionally important groundwater crustaceans. Effects of Carbo-Iron on H. azteca were determined in a 10-d acute test, a 7-d feeding activity test and a 42-d chronic test. Additionally, a 56-d life cycle test was performed with a modified design to further evaluate effects of Carbo-Iron on adult H. azteca and their offspring. The size of Carbo-Iron particles in stock and test suspensions was determined via dynamic light scattering. Potential uptake of particles into test organisms was investigated using transmission and scanning electron microscopy. At the termination of the feeding and acute toxicity test (i.e. after 7 and 10 d of exposure, respectively), Carbo-Iron had a significant effect on the weight, length and feeding rate of H. azteca at the highest test concentration of 100mg/L. While an uptake of Carbo-Iron into the gut was observed, no passage into the surrounding tissue was detected. In both chronic tests, the number of offspring was the most sensitive endpoint and significant effects were recorded at concentrations ≥50mg/L (42-d experiment) and ≥12.5mg/L (56-d experiment). Parental exposure to oxidized Carbo-Iron significantly exacerbated the acute effects of the nanocomposite on the subsequent generation of H. azteca by a factor >10. The present study indicates risks for groundwater species at concentrations in the mg/L range. Carbo-Iron may exceed these effect concentrations in treated aquifers, but the presence of the pollutant has most likely impaired the quality of this habitat already. The benefit of remediation has to be regarded against the risk of ecological consequences with special consideration of the observed increasing sensitivity of juvenile H. azteca.

Microplastics in the aquatic and terrestrial environment: sources (with a specific focus on personal care products), fate and effects.

Due to the widespread use and durability of synthetic polymers, plastic debris occurs in the environment worldwide. In the present work, information on sources and fate of microplastic particles in the aquatic and terrestrial environment, and on their uptake and effects, mainly in aquatic organisms, is reviewed. Microplastics in the environment originate from a variety of sources. Quantitative information on the relevance of these sources is generally lacking, but first estimates indicate that abrasion and fragmentation of larger plastic items and materials containing synthetic polymers are likely to be most relevant. Microplastics are ingested and, mostly, excreted rapidly by numerous aquatic organisms. So far, there is no clear evidence of bioaccumulation or biomagnification. In laboratory studies, the ingestion of large amounts of microplastics mainly led to a lower food uptake and, consequently, reduced energy reserves and effects on other physiological functions. Based on the evaluated data, the lowest microplastic concentrations affecting marine organisms exposed via water are much higher than levels measured in marine water. In lugworms exposed via sediment, effects were observed at microplastic levels that were higher than those in subtidal sediments but in the same range as maximum levels in beach sediments. Hydrophobic contaminants are enriched on microplastics, but the available experimental results and modelling approaches indicate that the transfer of sorbed pollutants by microplastics is not likely to contribute significantly to bioaccumulation of these pollutants. Prior to being able to comprehensively assess possible environmental risks caused by microplastics a number of knowledge gaps need to be filled. However, in view of the persistence of microplastics in the environment, the high concentrations measured at some environmental sites and the prospective of strongly increasing concentrations, the release of plastics into the environment should be reduced in a broad and global effort regardless of a proof of an environmental risk.

The oxidized state of the nanocomposite Carbo-Iron® causes no adverse effects on growth, survival and differential gene expression in zebrafish.

For degradation of halogenated chemicals in groundwater Carbo-Iron®, a composite of activated carbon and nano-sized Fe(0), was developed (Mackenzie et al., 2012). Potential effects of this nanocomposite on fish were assessed. Beyond the contaminated zone Fe(0) can be expected to have oxidized and Carbo-Iron was used in its oxidized form in ecotoxicological tests. Potential effects of Carbo Iron in zebrafish (Danio rerio) were investigated using a 48 h embryo toxicity test under static conditions, a 96 h acute test with adult fish under semi-static conditions and a 34 d fish early life stage test (FELST) in a flow-through system. Particle diameters in test suspensions were determined via dynamic light scattering (DLS) and ranged from 266 to 497 nm. Particle concentrations were measured weekly in samples from the FELST using a method based on the count rate in DLS. Additionally, uptake of particles into test organisms was investigated using microscopic methods. Furthermore, effects of Carbo-Iron on gene expression were investigated by microarray analysis in zebrafish embryos. In all tests performed, no significant lethal effects were observed. Furthermore, Carbo-Iron had no significant influence on weight and length of fish as determined in the FELST. In the embryo test and the early life stage test, growth of fungi on the chorion was observed at Carbo-Iron concentrations between 6.3 and 25mg/L. Fungal growth did not affect survival, hatching success and growth. In the embryo test, no passage of Carbo-Iron particles into the perivitelline space or the embryo was observed. In juvenile and adult fish, Carbo-Iron was detected in the gut at the end of exposure. In juvenile fish exposed to Carbo-Iron for 29 d and subsequently kept for 5d in control water, Carbo-Iron was no longer detectable in the gut. Global gene expression in zebrafish embryos was not significantly influenced by Carbo-Iron.

The zebrafish embryo model in toxicology and teratology, September 2­3, 2010, Karlsruhe, Germany.

The use of fish embryos is gaining popularity for research in the area of toxicology and teratology. Particularly embryos of the zebrafish offer an array of different applications ranging from regulatory testing to mechanistic research. For this reason a consortium of two research centres and a company with the support of the COST Action EuFishBiomed has organised the Workshop "The zebrafish embryo model in toxicology and teratology", in Karlsruhe, Germany, 2nd­3rd September 2010. The workshop aimed at bringing together experts from different areas of toxicology using the (zebra)fish embryo and stimulating networking between scientists and representatives from regulatory bodies, research institutions and industry. Recent findings, presented in various platform presentations in the area of regulatory toxicity, high throughput screening, toxicogenomics, as well as environmental and human risk assessment are highlighted in this meeting report. Furthermore, the constraints and possibilities of the model as discussed at the workshop are described. A follow up-meeting was appreciated by the about 120 participants and is planned for 2012.

Environmental risk assessment of ivermectin: A case study.

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.

Effects of ivermectin-spiked cattle dung on a water-sediment system with the aquatic invertebrates Daphnia magna and Chironomus riparius.

A two-species test using a water-sediment test system was performed to investigate chronic effects of the parasiticide ivermectin on Daphnia magna and Chironomus riparius. To simulate exposure by direct excretion of cattle into surface waters, ivermectin was applied via spiked cattle dung. The parasiticide was applied once, at concentrations ranging from 11 to 1314 microg kg(-1) dung dry weight. The highest concentration corresponds to the maximum concentration in dung 3 days after topical application to cattle. Test vessels were stocked with chironomid larvae and daphnids of defined, mixed age. Replicates were sampled 10, 24, 38 and 51 days post application. Survival, growth and emergence of chironomids, and abundance and biomass of daphnids were evaluated. In case of extinction of the D. magna population in all replicates of a concentration level, daphnids were re-introduced into the remaining vessels of this concentration to simulate immigration. In addition, a second batch of chironomid larvae was introduced into the vessels on day 27 post application. At 1314 microg ivermectin kg(-1) dung dry weight, survival, larval growth and emergence of the initially stocked chironomids were strongly affected. A significant effect on emergence was also observed for the second batch of chironomids. The two highest test concentrations led to 100% mortality of the initially stocked daphnids. At 1314 microg kg(-1) dung dry weight, no daphnids survived following re-introduction on days 11, 28 and 42. At 263 microg kg(-1) dung dry weight, the daphnids that were re-introduced on day 11 survived and reproduced, but abundance and biomass were reduced. The results of the present study indicate that following single application, toxic ivermectin concentrations persisted for an extended period. Possible effects on aquatic invertebrates, which may be caused by direct excretion of ivermectin-containing dung into surface water, deserve further attention.

Recommendations on the environmental risk assessment of pharmaceuticals: Effect characterization.

The effects testing of pharmaceuticals consists of a tiered investigation of ecotoxicological endpoints. However, effects testing has to be performed only when the predicted environmental concentrations (PECs) of pharmaceuticals are above certain action limits. To study the appropriateness of these action limits, a literature search was performed for pharmaceuticals with predicted no-effect concentrations (PNECs) close to or below the action limits. Some human pharmaceuticals showed effects at concentrations ≤100 ng/L, mostly in nonstandard fish or invertebrate tests. In addition, antibiotics and parasiticides sometimes had effects at concentrations <10 mg/kg soil. To help in identifying pharmaceuticals that should undergo effects testing although their PECs are below the action limits, "however clauses" are postulated for pharmaceuticals that are potentially persistent, bioaccumulative, carcinogenic, mutagenic, or reproductively toxic. Effects testing should also be performed for pharmaceuticals that 1) affect target structures that are conserved across species, 2) have a high potency or a small therapeutic margin, 3) are from a new therapeutic class, and 4) are structurally similar to compounds with known effects. Furthermore, suggestions for improving the effects testing of pharmaceuticals are made. These include inter alia chronic effects testing as a general approach, the use of invertebrate tests including sexual reproduction, the application of endpoints reflecting the mode of action of the drug or known side effects, and the simulation of more realistic exposure conditions in terrestrial laboratory tests.

Environmental risk assessment of human pharmaceuticals in the European Union: A case study with the ß-blocker atenolol.

ß-Adrenergic receptor blockers (ß-blockers) are applied to treat high blood pressure, ischemic heart disease, and heart rhythm disturbances. Due to their widespread use and limited human metabolism, ß-blockers are widely detected in sewage effluents and surface waters. ß-Adrenergic receptors have been characterized in fish and other aquatic animals, so it can be expected that physiological processes regulated by these receptors in wild animals may be affected by the presence of ß-blockers. Because ecotoxicological data on ß-blockers are scarce, it was decided to choose the ß-blocker atenolol as a case study pharmaceutical within the project ERAPharm. A starting point for the assessment of potential environmental risks was the European guideline on the environmental risk assessment of medicinal products for human use. In Phase I of the risk assessment, the initial predicted environmental concentration (PEC) of atenolol in surface water (500 ng L−1) exceeded the action limit of 10 ng L−1. Thus, a Phase II risk assessment was conducted showing acceptable risks for surface water, for groundwater, and for aquatic microorganisms. Furthermore, atenolol showed a low potential for bioaccumulation as indicated by its low lipophilicity (log KOW = 0.16), a low potential for exposure of the terrestrial compartment via sludge (log KOC = 2.17), and a low affinity for sorption to the sediment. Thus, the risk assessment according to Phase II-Tier A did not reveal any unacceptable risk for atenolol. Beyond the requirements of the guideline, additional data on effects and fate were generated within ERAPharm. A 2-generation reproduction test with the waterflea Daphnia magna resulted in the most sensitive no-observed-effect concentration (NOEC) of 1.8 mg L−1. However, even with this NOEC, a risk quotient of 0.003 was calculated, which is still well below the risk threshold limit of 1. Additional studies confirm the outcome of the environmental risk assessment according to EMEA/CHMP (2006). However, atenolol should not be considered as representative for other ß-blockers, such as metoprolol, oxprenolol, and propranolol, some of which show significantly different physicochemical characteristics and varying toxicological profiles in mammalian studies.

Gene expression analysis in zebrafish embryos: a potential approach to predict effect concentrations in the fish early life stage test.

Based on the hypothesis that analysis of gene expression could be used to predict chronic fish toxicity, the zebrafish (Danio rerio) embryo test (DarT), developed as a replacement method for the acute fish test, was expanded to a gene expression D. rerio embryo test (Gene-DarT). The effects of 14 substances on lethal and sublethal endpoints of the DarT and on expression of potential marker genes were investigated: the aryl hydrocarbon receptor 2, cytochrome P450 1A (cypla), heat shock protein 70, fizzy-related protein 1, the transcription factors v-maf musculoaponeurotic fibrosarcoma oncogene family protein g (avian) 1 and NF-E2-p45-related factor, and heme oxygenase 1 (hmox1). After exposure of zebrafish embryos for 48 h, differential gene expression was evaluated using reverse transcriptase-polymerase chain reaction, gel electrophoresis, and densitometric analysis of the gels. All tested compounds significantly affected the expression of at least one potential marker gene, with cyp1a and hmox1 being most sensitive. Lowest-observed-effect concentrations (LOECs) for gene expression were below concentrations resulting in 10% lethal effects in the DarT. For 10 (3,4- and 3,5-dichloroaniline, 1,4-dichlorobenzene, 2,4-dinitrophenol, atrazine, parathion-ethyl, chlorotoluron, genistein, 4-nitroquinoline-1-oxide, and cadmium) out of the 14 tested substances, LOEC values derived with the Gene-DarT differ by a factor of less than 10 from LOEC values of fish early life stage tests with zebrafish. For pentachloroaniline and pentachlorobenzene, the Gene-DarT showed a 23- and 153-fold higher sensitivity, respectively, while for lindane, it showed a 13-fold lower sensitivity. For ivermectin, the Gene-DarT was by a factor of more than 1,000 less sensitive than the acute fish test. The results of the present study indicate that gene expression analysis in zebrafish embryos could principally be used to predict effect concentrations in the fish early life stage test.

Effects of the parasiticide ivermectin on the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata.

Although widely used for the treatment of endo- and ectoparasites in livestock and pets, very few data on chronic effects on aquatic organisms are available for the parasiticide ivermectin. In the present study, toxicity of ivermectin to two freshwater organisms, the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata was investigated. For D. magna, a mean LC(50) 48 h of 5.7 ngl(-1) was derived from 10 acute tests. Chronic toxicity of ivermectin to D. magna was extremely high: with 0.001 and 0.0003 ngl(-1), respectively, nominal LOEC and NOEC based on growth and reproduction were far below the analytical limit of detection for this compound. P. subcapitata was considerably less sensitive to ivermectin than D. magna. For both growth rate and yield, EC(50) was >4,000 microgl(-1), LOEC was 1,250 microgl(-1) and NOEC 391microgl(-1). In view of the high toxicity to D. magna, the use of ivermectin might pose a risk to local aquatic ecosystems. Further studies should be carried out to investigate the effects of ivermectin and its degradation products on pelagic and benthic freshwater invertebrates.

The EU-project ERAPharm. Incentives for the further development of guidance documents?

Triggered by the detection of a large variety of pharmaceuticals in surface waters, soils and groundwaters across the world (e.g. Halling-Sørensen et al. 1998, Daughton & Ternes 1999, Jones et al. 2001, Heberer 2002) and the widespread occurrence of endocrine active compounds and related effects in the environment (e.g. Purdom et al. 1994, Tyler et al. 1998, Vethaak et al. 2002), pharmaceuticals in the environment have become an issue for both the scientific and the public community. During the last few years, our understanding of the fate and effects of pharmaceuticals in the environment has progressed significantly. However, there are still a number of uncertainties concerning the effects of pharmaceuticals on the environment and the assessment of potential exposure (e.g. Hanisch et al. 2004, Salomon 2005). These uncertainties will be addressed by the EU-project 'Environmental risk assessment of pharmaceuticals' (ERAPharm). This project, a specific targeted research project, is carried out within the priority 'Global change and ecosystems' of the 6th framework programme of the European Union. ERAPharm has started on 1st October 2004; the project duration is three years.