Bioassay predictive values for chemical health risks in drinking water.

Bioanalytical tools can be used for assessment of the chemical quality of drinking water and its sources. For water managers it is important to know the probability that a bioassay response above an established health-based 'effect-based trigger value' (EBT) indeed implies a harmful chemical (mixture) concentration. This study presents and applies a framework, based on Bayes' theorem, to derive such risk probabilities for bioassay responses. These were evaluated under varying (in silico) chemical mixture concentrations relevant to drinking water (sources), with toxicity data for six in vitro assays from the ToxCast database. For single chemicals and in silico mixtures, the negative predictive value (NPV) was 100 % for all assays. For water managers, this means that when a bioassay response is below the EBT, a chemical risk is reliably absent, and no further action is required. The positive predictive value (PPV) increased with increasing chemical concentrations (2 µg/L) up to 40-80 %, depending on the assay. For in silico mixtures of increasing numbers of chemicals, the PPV did not increase until higher sum concentrations (>2-10 µg/L). Hence, the ability to accurately signal a harmful chemical (mixture) using bioassays will be lowest for highly diverse, low-concentration chemical mixtures. For water managers, this means in practice that further investigations after an EBT exceedance will, in many cases, not reveal chemicals at harmful concentrations. A solution offered is to increase the trigger value for positive responses to achieve a higher PPV and maintain the EBT for negative responses to ensure an optimal NPV.

European-wide spatial analysis of sewage treatment plants and the possible benefits to nature of advanced treatment to reduce pharmaceutical emissions.

Pharmaceuticals are known to widely occur in the environment and to affect the health of ecosystems. Sewage treatment plants (STPs) are main emission pathways for pharmaceuticals, which are often not sufficiently removed during wastewater treatment. In Europe, STP treatment requirements are specified under the Urban WasteWater Treatment Directive (UWWTD). The introduction of advanced treatment techniques, such as ozonation and activated carbon, under the UWWTD is expected to be an important option to reduce pharmaceutical emissions. In this study, we present a European-wide analysis of STPs reported under the UWWTD, their current treatment level and potential to remove a set of 58 prioritised pharmaceuticals. Three different scenarios were analysed to show 1) UWWTD present effectiveness, 2) the effectiveness at full UWWTD compliance, and 3) the effectiveness when advanced treatment is implemented at STPs with a treatment capacity of >100.000 person equivalents. Based on a literature study, the potential of individual STPs to reduce pharmaceutical emissions ranged from an average of 9% for STPs with primary treatment to 84% for STPs applying advanced treatment. Results of our calculations show that European-wide emission of pharmaceuticals can be reduced with 68% when large STPs are updated with advanced treatment, but spatial differences exist. We argue that adequate attention should also be paid with regards to preventing environmental impacts of STPs with a capacity <100.000 p.e. Circa 44% of total STP effluent is emitted near Natura2000 sites (EU nature protection areas). Of all surface waters receiving STP effluent for which the ecological status has been assessed under the Water Framework Directive, 77% have a status of less than good. Relatively often only primary treatment is applied to wastewater emitted into coastal waters. This analysis can be used to further model pharmaceutical concentrations in European surface waters, to identify STPs for which more advanced treatment might be required and to protect EU aquatic biodiversity.

Microplastic discharge from a wastewater treatment plant: long term monitoring to compare two analytical techniques, LDIR and optical microscopy while also assessing the removal efficiency of a bubble curtain.

In this study we compare two parallel analytical methods while also testing a microplastics mitigation method. We assess the effectiveness of a bubble curtain to reduce microplastics in a wastewater treatment plant (WWTP)-effluent canal during the course of six months (>70 samples) using two analytical techniques: laser direct infrared (LDIR) and optical microscopy (OM) covering a size range of 0.02 to 5 mm. Comparison of the two analytical strategies shows similar trends, fluctuations, and correlating particle and fibre numbers. However, absolute values of particles differ, and the strategies provide different levels of information: LDIR is capable of identifying the plastic type as well as shape, while OM cannot determine the plastic type. Furthermore LIDR has a lower size limit (10-20 µm) than OM (50 µm). While information obtained by OM in general is far less detailed it is more affordable. This research also shows that the bubble curtain pilot does not have a measurable effect on the particle concentration. Possible effects of the curtain are hidden in the temporal variations. This research also reveals that individual samples show a large variation in particle numbers, illustrating that single measurements might give a poor representation of environmental particle number.

Effect-based monitoring to integrate the mixture hazards of chemicals into water safety plans.

Water safety plans (WSPs) are intended to assure safe drinking water (DW). WSPs involve assessing and managing risks associated with microbial, chemical, physical and radiological hazards from the catchment to the consumer. Currently, chemical hazards in WSPs are assessed by targeted chemical analysis, but this approach fails to account for the mixture effects of the many chemicals potentially present in water supplies and omits the possible effects of non-targeted chemicals. Consequently, effect-based monitoring (EBM) using in vitro bioassays and well plate-based in vivo assays are proposed as a complementary tool to targeted chemical analysis to support risk analysis, risk management and water quality verification within the WSP framework. EBM is frequently applied to DW and surface water and can be utilised in all defined monitoring categories within the WSP framework (including 'system assessment', 'validation', 'operational' and 'verification'). Examples of how EBM can be applied within the different WSP modules are provided, along with guidance on where to apply EBM and how frequently. Since this is a new area, guidance documents, standard operating procedures (SOPs) and decision-making frameworks are required for both bioassay operators and WSP teams to facilitate the integration of EBM into WSPs, with these resources being developed currently.

Comparing conventional and green fracturing fluids by chemical characterisation and effect-based screening.

There is public and scientific concern about air, soil and water contamination and possible adverse environmental and human health effects as a result of hydraulic fracturing activities. The use of greener chemicals in fracturing fluid aims to mitigate these effects. This study compares fracturing fluids marketed as either 'conventional' or 'green', as assessed by their chemical composition and their toxicity in bioassays. Chemical composition was analysed via non-target screening using liquid chromatography - high resolution mass spectrometry, while toxicity was evaluated by the Ames fluctuation test to assess mutagenicity and CALUX reporter gene assays to determine specific toxicity. Overall, the results do not indicate that the 'green' fluids are less harmful than the 'conventional' ones. First, there is no clear indication that the selected green fluids contain chemicals present at lower concentrations than the selected conventional fluids. Second, the predicted environmental fate of the identified compounds does not seem to be clearly distinct between the 'green' and 'conventional' fluids, based on the available data for the top five chemicals based on signal intensity that were tentatively identified. Furthermore, Ames fluctuation test results indicate that the green fluids have a similar genotoxic potential than the conventional fluids. Results of the CALUX reporter gene assays add to the evidence that there is no clear difference between the green and conventional fluids. These results do not support the claim that currently available and tested green-labeled fracturing fluids are environmentally more friendly alternatives to conventional fracturing fluids.

Organic Pollutants in Shale Gas Flowback and Produced Waters: Identification, Potential Ecological Impact, and Implications for Treatment Strategies.

Organic contaminants in shale gas flowback and produced water (FPW) are traditionally expressed as total organic carbon (TOC) or chemical oxygen demand (COD), though these parameters do not provide information on the toxicity and environmental fate of individual components. This review addresses identification of individual organic contaminants in FPW, and stresses the gaps in the knowledge on FPW composition that exist so far. Furthermore, the risk quotient approach was applied to predict the toxicity of the quantified organic compounds for fresh water organisms in recipient surface waters. This resulted in an identification of a number of FPW related organic compounds that are potentially harmful namely those compounds originating from shale formations (e.g., polycyclic aromatic hydrocarbons, phthalates), fracturing fluids (e.g., quaternary ammonium biocides, 2-butoxyethanol) and downhole transformations of organic compounds (e.g., carbon disulfide, halogenated organic compounds). Removal of these compounds by FPW treatment processes is reviewed and potential and efficient abatement strategies are defined.

Release of primary microplastics from consumer products to wastewater in the Netherlands.

The authors estimate the release of primary microplastics from consumer products-cosmetics and personal care products, cleaning agents, and paint and coatings-via sewage effluent as an expected relevant route to the marine environment. Total estimated concentrations in the 3 scenarios are 0.2 µg/L, 2.7 µg/L, and 66 µg/L in sewage-treatment plant (STP) effluent, respectively. All product categories relevantly contribute. Predicted concentrations are compared with reported actual concentrations in STP effluents. Environ Toxicol Chem 2016;35:1627-1631. © 2015 SETAC.

Daphnid life cycle responses to the insecticide chlorantraniliprole and its transformation products.

Chlorantraniliprole (CAP) is a newly developed, widely applied insecticide. In the aquatic environment, several transformation products are formed under natural conditions, one by dehydration and others by photoinduced degradation. Data on aquatic ecotoxicity of CAP can mainly be found in registration and regulatory evaluation reports. Moreover, the toxicity of its transformation products and especially effects upon chronic exposure remain completely unknown. Hence, our aim was to investigate the acute and chronic toxicity of CAP and its transformation products to the daphnid Daphnia magna. The results showed that CAP is extremely toxic to D. magna, with an acute and chronic LC50 of 9.4 and 3.7 µg/L, respectively. No effects on daphnid reproduction were observed, but the impact on daphnid survival also affected population growth rate, with an EC50 of 3.5 µg/L. In contrast, no negative effects of the two main degradation products were observed. The present study demonstrated a high sensitivity of nontarget microcrustaceans to CAP. However, the actual risk of CAP in water diminishes with its spontaneous or light-induced degradation into two transformation products, showing no toxicity to the daphnids in the present study.

Daphnid life cycle responses to new generation flame retardants.

Relatively hazardous brominated flame retardants (BFRs) are currently substituted with halogen-free flame retardants (HFFRs). Consequently, information on their persistence, bioaccumulation and toxicity (PBT) is urgently needed. Therefore, we investigated the chronic toxicity to the water flea Daphnia magna of two HFFRs, aluminum diethylphosphinate (ALPI) and 9,10-dihyro-9-oxa-10-phosphaphenanthrene-oxide (DOPO). The toxicity of ALPI increased from a 48 h LC50 of 18 mg L(-1) to a 21 day LC50 value of 3.2 mg L(-1), resulting in an acute-to-chronic ratio of 5.6. This may imply a change in classification from low to moderate toxicity. ALPI also affected sublethal life cycle parameters, with an EC50 of 2.8 mg L(-1) for cumulative reproductive output and of 3.4 mg L(-1) for population growth rate, revealing a nonspecific mode of action. DOPO showed only sublethal effects with an EC50 value of 48 mg L(-1) for cumulative reproductive output and an EC50 value of 73 mg L(-1) for population growth rate. The toxicity of DOPO to D. magna was classified as low and likely occurred above environmentally relevant concentrations, but we identified specific effects on reproduction. Given the low chronic toxicity of DOPO and the moderate toxicity of ALPI, based on this study only, DOPO seems to be more suitable than ALPI for BFR replacement in polymers.

Toxicity of new generation flame retardants to Daphnia magna.

There is a tendency to substitute frequently used, but relatively hazardous brominated flame retardants (BFRs) with halogen-free flame retardants (HFFRs). Consequently, information on the persistence, bioaccumulation and toxicity (PBT) of these HFFRs is urgently needed, but large data gaps and inconsistencies exist. Therefore, in the present study the toxicity of a wide range of HFFRs to the water flea Daphnia magna was investigated. Our results revealed that four HFFRs were showing no effect at their Sw (saturated water concentration) and three had a low toxicity (EC50>10 mg L(-1)), suggesting that these compounds are not hazardous. Antimony trioxide had a moderate toxicity (EC50=3.01 mg L(-1), 95% CL: 2.76-3.25) and triphenyl phosphate and the brominated reference compound tetra bromobisphenol A were highly toxic to D. magna (EC50=0.55 mg L(-1), 95% CL: 0.53-0.55 and EC50=0.60 mg L(-1), 95% CL: 0.24-0.97 respectively). Aluminum trihydroxide and bisphenol A bis(diphenyl phosphate) caused limited mortality at Sw (26 and 25% respectively) and have a low solubility (<10 mg L(-1)). Hence, increased toxicity of these compounds may be observed when for instance decreasing pH could increase solubility. By testing all compounds under identical conditions we provided missing insights in the environmental hazards of new generation flame retardants and propose as best candidates for BFR replacements: APP, ALPI, DOPO, MHO, MPP, ZHS and ZS.

Assessment of structure and function in metal polluted grasslands using Terrestrial Model Ecosystems.

Ecosystem effects of metal pollution in field situations are hard to predict, since metals occur often in mixtures and links between structural (organisms) and functional endpoints (ecosystem processes) are not always that clear. In grasslands, both structure and functioning was suspected to be affected by a mixture of copper, lead, and zinc. Therefore, the structural and functional variables were studied simultaneously using Terrestrial Model Ecosystems (TMEs). Comparing averages of low- and high-polluted soil, based on total metal concentrations, did not show differences in structural and functional variables. However, nematode community structure (Maturity Index) negatively correlated with metal concentrations. Next to that, multivariate statistics showed that enchytraeid, earthworm and, to lesser extent, nematode diversity decreased with increasing metal concentrations and a lower pH in the soil. Bacterial CFU and nematode biomass were positively related with decomposer activity and nitrate concentrations. Nitrate concentrations were negatively related to ammonium concentrations. Earthworm biomass, CO(2) production and plant yield were not related to metal concentrations. The most metal-sensitive endpoint was enchytraeid biomass. In all analyses, soil pH was a significant factor, indicating direct effects on organisms, or indicating indirect effects by influencing metal availability. In general, structural diversity seemed more positively related to functional endpoints than structural biomass. TMEs proved valuable tools to assess the structure and function in metal polluted field situations. The outcome feeds modeling effort and direct future research.

Stress responses investigated; application of zinc and heat to Terrestrial Model Ecosystems from heavy metal polluted grassland.

This study tested the hypothesis that soils with a deprived biodiversity due to metal pollution are less stable than non-polluted soils, containing a more diverse community. For this, soils were sampled from specific grasslands in the Netherlands that contain elevated heavy metal concentrations (Cu, Pb and Zn). Soils that showed the largest differences in metal concentrations were incubated in the laboratory using Terrestrial Model Ecosystems (TMEs). This approach enabled simultaneous measurement of structural (bacteria, nematodes, enchytraeids, earthworms) and functional parameters (nitrogen leaching, feeding activity, CO2 production, plant growth). The highest polluted soils showed a lower bacterial growth, and decreased enchytraeid and nematode biomass and diversity, hence a deprived community. More nitrate leached from high polluted soils, while all other functional endpoints did not differ. Additional stress application of zinc and heat was used to test the stability. Zinc treatment caused effects only in the higher polluted soils, observed at several moments in time for enchytraeids, CO2 fluxes and plant growth. Heat stress caused a large reduction in enchytraeid and earthworm biomass. Ammonium leaching was decreased by heat treatments in the most polluted soils, while CO2 was increased by heat in less polluted soils. Most effects were seen in the most polluted systems and it was concluded that they seem less stable.

A ranking of European veterinary medicines based on environmental risks.

The most likely entry pathways of veterinary pharmaceuticals to the environment are via slurry or manure from intensively reared animals to soil and via dung or urine from animals grazing on pasture. These pathways may result in contamination of surface water via runoff or leaching and drainage. Direct entry into water may occur by defecation by pasture animals or by Scompanion animals. In addition, application of medicines for aquaculture is important for a limited number of veterinary medicinal products. For a large number of veterinary medicinal products, consistent data on the environmental risk have never been generated. In this project, a simple risk-based ranking procedure was developed that should allow assessing the potential for environmental risks of active substances of veterinary medicinal products. In the European Union approximately 2000 products containing 741 active substances were identified. In the prescreening step and in agreement with the technical guidelines released by the European Medicines Agency, 294 natural substances, complex mixtures, and substances with low expected exposure were exempted from the ranking procedure. For 233 active substances, sufficient information was collated on 4 exposure scenarios: Intensively reared animals, pasture animals, companion animals, and aquaculture. The ranking approach was performed in 4 phases: (1) usage estimation; (2) characterization of exposure to soil, dung, surface water, and aquatic organisms depending on exposure scenarios; (3) characterization of effects based on therapeutical doses; and (4) risk characterization, which is the ratio of exposure to effects (risk index), and ranking. Generally, the top-ranked substances were from the antibiotic and parasiticide groups of veterinary medicines. Differences occurred in the ranking of substances in soil via application to either intensively reared or pasture animals. In intensive rearing, anticoccidia, for example, are used as feed-administered medicines (feed additives) in comparatively large doses over a long time. For pasture animals, these substances are used less, if at all, and therefore receive lower ranks. Besides that, the risk indices for the aquatic compartment are large for substances used in aquaculture or applied to companion animals. In conclusion, the ranking scheme developed for this project provided a scientifically based and pragmatic means of assessing the relative priority of veterinary medicines for further detailed risk assessment. The outcome of this project will support pharmaceutical industries and competent authorities when seeking authorization for market applications of veterinary pharmaceutical products.

Estimating the use of veterinary medicines in the European union.

Risk-based ranking is a procedure that ranks active substances according to their potential environmental risk, which can be used to prioritise effort in environmental risk assessment procedures. An important aspect in this procedure is the extent of use of the active substance. However, use volume data for veterinary pharmaceuticals are generally not publicly available. Therefore, the use of several groups of pharmaceuticals was estimated. For this, available data were collected from different sources and used to calculate the average use, related with the total meat production in the respective countries. Then, an extrapolation, based on food-stuff production data, yields a rough estimate of the use volume for all European countries, largely based on 2004 data. The estimated use was 5393 tons antibiotics, 194 tons antiparasitics and 4.6 tons hormones, 221 tons to treat the alimentary tract and metabolism related disorders, 120 tons CNS active substances, 60 tons of substances used for blood and blood forming organs and 52 tons for muscles and skeleton related disorders. In total, an estimated 6051 tons of active substances in veterinary medicines were used in the European Union. We discuss available data sources and quality, our method and possible refinement of our approach. Environmental risk-based ranking procedures of veterinary substances would benefit from more detailed use data.

Microbial responses to zinc in soil microcosms with and without a natural assemblage of enchytraeids.

The presence of higher trophic levels in studies on the toxicity on soil contaminants to microbial processes increases ecological realism. This study assessed the toxicity of zinc to soil microbial processes in the presence and absence of enchytraeids (Oligochaeta, Annelida). We incubated microcosms under standard conditions without or inoculated with a natural assemblage of enchytraeid species. Total zinc concentrations of 365 to 1,360 mg/kg caused no mortality of enchytraeids during six weeks' incubation. Soil nitrate concentrations showed a negative trend under zinc addition and soil ammonium concentrations were the highest at zinc concentrations of 1,360 mg/kg, indicating impairment of ammonium oxidation. Zinc decreased bacterial carbon biomass and caused a dose-response decrease of the respiration, but this was not observed in the presence of enchytraeids. Respiration, ammonium concentrations, and soil moisture contents were increased by enchytraeids. We observed no interaction between the addition of zinc and the presence of enchytraeids. The effect of enchytraeids on soil-moisture contents and microbial processes, and the importance of enchytraeid-microbial interactions are discussed.

Variation in sex steroids and phallus size in juvenile American alligators (Alligator mississippiensis) collected from 3 sites within the Kissimmee-Everglades drainage in Florida (USA).

This 3-year study was designed to examine variation in plasma sex steroids, phallus size, and the standard error (S.E.) associated with these endpoints in juvenile alligators collected from 3 sites within the Kissimmee-Everglades drainage (Florida, USA) with varying concentrations of sediment organochlorine contaminants. We hypothesized that decreased plasma sex steroid concentrations and phallus size would be observed in the higher contaminant site when compared to the intermediate and lower contaminant sites. Furthermore, we hypothesized that greater S.E. associated with these endpoints would be observed for the populations from more contaminated sites. We found that differences existed with females from the higher contaminant site exhibiting lower plasma estradiol-17beta (E2) and testosterone (T) concentrations. Males from the higher contaminant site exhibited smaller phallus sizes than males from the intermediate and lower contaminant sites. Smaller phallus size in this case differed from that reported in Lake Apopka male alligators [Gen. Comp. Endocrinol. 116 (1999) 356] in that a significant positive relationship between body size and phallus size existed. No difference among sites was observed in plasma T for males. Lower S.E. was associated with E2 and T concentrations in females from the higher contaminant site and in phallus size in males from the higher contaminant site. This pattern was opposite to what we had hypothesized. We concluded that variation in plasma E2 and T concentrations, phallus size, and the S.E. associated with these endpoints exists among the 3 sites with the patterns matching the patterns of organochlorine contamination, although S.E. patterns were opposite to what was predicted.

Temporal and spatial variation in plasma thyroxine (T4) concentrations in juvenile alligators collected from Lake Okeechobee and the northern Everglades, Florida, USA.

We examined variation in plasma thyroxine (T4) in juvenile American alligators (Alligator mississippiensis) collected from three sites within the Kissimmee River drainage basin (FL, USA). Based on historical sediment data, Moonshine Bay served as the low contaminant exposure site, Water Conservation Area 3A served as an intermediate contaminant exposure site, and Belle Glade served as the high contaminate exposure site. In May 1999, alligators (n = 22) from Water Conservation Area 3A exhibited higher T4 concentrations than animals from both Belle Glade (n = 22; p = 0.0003) and Moonshine Bay (n = 33; p = 0.001). In May 2000, alligators (n = 29) Water Conservation Area 3A again exhibited higher T4 concentrations than those from Belle Glade (n = 49; p = 0.02) but not those from Moonshine Bay (n = 40). No sexual dimorphism was observed among mean T4 concentrations within any of the sites during either year (p > 0.05). Animals within all sites exhibited higher T4 concentrations in May 2000 when compared to May 1999. When variance was examined, animals from Water Conservation Area 3A exhibited higher variance in plasma T4 concentrations than those from either Moonshine Bay or Belle Glade. We concluded that mean plasma T4 concentrations did not match the sediment contaminant mixture data presently available to us, whereas variance seems to be a more reliable indicator of contaminant exposure.