Exploring BPA alternatives - Environmental levels and toxicity review.

Bisphenol A alternatives are manufactured as potentially less harmful substitutes of bisphenol A (BPA) that offer similar functionality. These alternatives are already in the market, entering the environment and thus raising ecological concerns. However, it can be expected that levels of BPA alternatives will dominate in the future, they are limited information on their environmental safety. The EU PARC project highlights BPA alternatives as priority chemicals and consolidates information on BPA alternatives, with a focus on environmental relevance and on the identification of the research gaps. The review highlighted aspects and future perspectives. In brief, an extension of environmental monitoring is crucial, extending it to cover BPA alternatives to track their levels and facilitate the timely implementation of mitigation measures. The biological activity has been studied for BPA alternatives, but in a non-systematic way and prioritized a limited number of chemicals. For several BPA alternatives, the data has already provided substantial evidence regarding their potential harm to the environment. We stress the importance of conducting more comprehensive assessments that go beyond the traditional reproductive studies and focus on overlooked relevant endpoints. Future research should also consider mixture effects, realistic environmental concentrations, and the long-term consequences on biota and ecosystems.

Ciona spp. and ascidians as bioindicator organisms for evaluating effects of endocrine disrupting chemicals: A discussion paper.

In context of testing, screening and monitoring of endocrine-disrupting (ED) type of environmental pollutants, tunicates could possibly represent a particularly interesting group of bioindicator organisms. These primitive chordates are already important model organisms within developmental and genomics research due to their central position in evolution and close relationship to vertebrates. The solitary ascidians, such as the genus Ciona spp. (vase tunicates), could possibly be extra feasible as ED bioindicators. They have a free-swimming, tadpole-like larval stage that develops extremely quickly (<20 h under favorable conditions), has a short life cycle (typically 2-3 months), are relatively easy to maintain in laboratory culture, have fully sequenced genomes, and transgenic embryos with 3D course data of the embryo ontogeny are available. In this article, we discuss possible roles of Ciona spp. (and other solitary ascidians) as ecotoxicological bioindicator organisms in general but perhaps especially for effect studies of contaminants with presumed endocrine disrupting modes of action.

The ecotoxicology of marine tributyltin (TBT) hotspots: A review.

Tributyltin (TBT) was widely used as a highly efficient biocide in antifouling paints for ship and boat hulls. Eventually, TBT containing paints became globally banned when TBT was found to cause widespread contamination and non-target adverse effects in sensitive species, with induced pseudohermaphroditism in female neogastropods (imposex) being the best-known example. In this review, we address the history and the status of knowledge regarding TBT pollution and marine TBT hotspots, with a special emphasis on the Norwegian coastline. The review also presents a brief update on knowledge of TBT toxicity in various marine species and humans, highlighting the current understanding of toxicity mechanisms relevant for causing endocrine disruption in marine species. Despite observations of reduced TBT sediment concentrations in many marine sediments over the recent decades, contaminant hotspots are still prevalent worldwide. Consequently, efforts to monitor TBT levels and assessment of potential effects in sentinel species being potentially susceptible to TBT in these locations are still highly warranted.

Alkylphenol metabolites in fish bile as biomarkers of exposure to offshore oil industry produced water in feral fish.

The measurement of low-concentration alkylphenol (AP) exposure in fish is relevant in connection with monitoring and risk assessment of offshore oil industry produced water (PW) discharges. Detection of AP markers in fish bile offers significantly greater sensitivity than detection of AP in tissues such as liver. Recent studies revealed that gas chromatography-mass spectrometry in electron ionization mode (GC-EI-MS) enabled a selective and sensitive analytical detection of PW AP in mixtures with unknown composition. A procedure consisting of enzymatic deconjugation of metabolites in fish bile followed by derivatization with bis(trimethylsilyl)trifluoroacetamide and then separation and quantification of derivatized AP using GC-EI-MS is presented. The use of this procedure as a possible recommended approach for assessment and biomonitoring of AP contamination in fish populations living down-current from offshore oil production fields is presented.

Environmental effects of offshore produced water discharges: A review focused on the Norwegian continental shelf.

Produced water (PW), a large byproduct of offshore oil and gas extraction, is reinjected to formations or discharged to the sea after treatment. The discharges contain dispersed crude oil, polycyclic aromatic hydrocarbons (PAHs), alkylphenols (APs), metals, and many other constituents of environmental relevance. Risk-based regulation, greener offshore chemicals and improved cleaning systems have reduced environmental risks of PW discharges, but PW is still the largest operational source of oil pollution to the sea from the offshore petroleum industry. Monitoring surveys find detectable exposures in caged mussel and fish several km downstream from PW outfalls, but biomarkers indicate only mild acute effects in these sentinels. On the other hand, increased concentrations of DNA adducts are found repeatedly in benthic fish populations, especially in haddock. It is uncertain whether increased adducts could be a long-term effect of sediment contamination due to ongoing PW discharges, or earlier discharges of oil-containing drilling waste. Another concern is uncertainty regarding the possible effect of PW discharges in the sub-Arctic Southern Barents Sea. So far, research suggests that sub-arctic species are largely comparable to temperate species in their sensitivity to PW exposure. Larval deformities and cardiac toxicity in fish early life stages are among the biomarkers and adverse outcome pathways that currently receive much attention in PW effect research. Herein, we summarize the accumulated ecotoxicological knowledge of offshore PW discharges and highlight some key remaining knowledge needs.

Uptake and tissue distribution of C4-C7 alkylphenols in Atlantic cod (Gadus morhua): relevance for biomonitoring of produced water discharges from oil production.

The sensitivity of different tissues for assessment of chronic low-dose environmental exposure of fish to alkylphenols (APs) was investigated. We exposed Atlantic cod (Gadus morhua) in the laboratory to tritium labelled 4-tert-butylphenol, 4n-pentylphenol, 4n-hexylphenol, and 4n-heptylphenol via seawater (8 ng/l) and via contaminated feed (5 microg/kg fish per day). Measurements of different fish tissues during eight days of exposure and eight subsequent days of recovery revealed that APs administered via spiked seawater were readily taken up whereas the uptake was far less efficient when APs were administered in spiked feed. AP residues were mainly located in the bile fluid whereas the concentrations in liver were very low, indicating a rapid excretion and the liver-bile axis to be the major route of elimination. The biological half-life of APs in the exposed cod was short, between 10 and 20 h. Our study shows that in connection with biomonitoring of AP exposure in fish, assessment of AP metabolites in bile fluid is a more sensitive tool than detection of parent AP levels in liver or other internal tissues.

Levels and trends of tributyltin (TBT) and imposex in dogwhelk (Nucella lapillus) along the Norwegian coastline from 1991 to 2017.

The banning of organotin biocides, such as tributyltin (TBT), from use in marine antifouling paints is now leading to reproductive health recovery in marine gastropod populations all over the world. TBT induces so-called imposex (superimposition of male sexual characters onto females) in certain marine gastropods, such as the common dogwhelk Nucella lapillus. In this publication, the results of the Norwegian TBT and imposex monitoring in N. lapillus from the period 1991-2017 are presented. Significantly higher levels of TBT and imposex were measured in coastal areas close to shipping lanes along most of the coast prior to 2008 than afterwards. Levels started declining after restrictions were imposed on the use of TBT in all antifouling paint applications, with a total ban in 2008. In 2017, no sign of imposex was found in N. lapillus in any of the monitoring stations along the Norwegian coastline. Based on monitoring data shown herein, the importance of long-term biomonitoring and international chemical regulations, as well as the TBT and imposex story in general, are discussed.

Solid-phase analytical derivatization of alkylphenols in fish bile for gas chromatography-mass spectrometry analysis.

Solid-phase analytical derivatization (SPAD) with bis(trimethylsilyl)trifluoroacetamide (BSTFA) has successfully been used as a sample preparation method for determination of (APs) in fish bile treated with beta-glucuronidase and sulfatase. Derivatized APs were analysed by gas chromatography-mass spectrometry in the electron ionization mode (GC-EI-MS). Overall limits of detection (LODs) ranged from 5 to 18ng/g bile for 19 out of 21 investigated compounds. LODs were not determined for 4-methylphenol and 4-tert-octylphenol due to high background levels in control bile. Recoveries ranged from 83 to 109%. The analysed APs vary in degree of alkylation from methyl (C(1)) to nonyl (C(9)), and represent various pollution sources, including produced water (PW) discharge from the offshore oil industry. The applicability and sensitivity of the method has been demonstrated by analysis of bile taken from Atlantic cod (Gadus morhua L.) exposed to two dilutions of PW (1:500 and 1:1500) in a continuous flow system.

Characterization of alkylphenol metabolites in fish bile by enzymatic treatment and HPLC-fluorescence analysis.

Alkylphenol (AP) metabolites were characterized in the bile of Atlantic cod (Gadus morhua L.) after exposure to nine individual compounds (10mg/kg fish), 2-methylphenol (2-MP), 4-methylphenol (4-MP), 3,5-dimethylphenol (3,5-DMP), 2,4,6-trimethylphenol (2,4,6-TMP), 4-tert-butylphenol (4-t-BP), 4-tert-butyl-2-methylphenol (4-t-B-2-MP), 4-n-pentylphenol (4-n-PP), 4-n-hexylphenol (4-n-HexP) and 4-n-heptylphenol (4-n-HepP), and a mixture (total dose; 13.5 mg/kg fish) of the nine APs by inter-muscular injection. The degree of alkylation ranged from methyl (C1) to heptyl (C7) and represents the types of APs present in produced water. Fish bile was collected on day 4 and 16 (exposure groups 2-MP, 3,5-DMP, 2,4,6-TMP and 4-t-B-2-MP) following exposure. Characterization of major metabolites was accomplished by enzymatic de-conjugation and analysis by high performance liquid chromatography connected to a fluorescence detector (HPLC-F) acquiring at ex/em 222/306 nm. Two solid phase extraction (SPE) columns were evaluated for clean-up of samples prior to analysis. Independent of alkyl homologue, the glucuronide conjugated APs were the most abundant metabolites (73-100%), whereas sulfates, glucosides and unchanged compounds were excreted in amounts of 0-21%, 0-6.1% and 0-6.3%, respectively. The total concentration of measured metabolites in the bile, determined as their respective APs after de-conjugation, increased with increasing degree of alkylation (3.2+/-2.6 microg/g bile for 2-MP and 571+/-81 microg/g bile for 4-n-HepP) after exposure to an equal dose of AP. Comparison of metabolite concentrations in bile sampled 4 and 16 days after exposure, showed that the levels of 2-MP, 2,4,6-TMP and 4-t-B-2-MP were reduced by 55%, 30% and 45%, respectively whereas 3,5-DMP increased by 25% (not significant). This study suggests that analysis of de-conjugated metabolites in fish bile can be used to monitor AP exposure to fish, due to the relatively high and persistent level of these compounds. However, although HPLC-F is suitable for laboratory exposures, it might not be sufficient selective for field studies.

Mytilus spp. as sentinels for monitoring microplastic pollution in Norwegian coastal waters: A qualitative and quantitative study.

Microplastic (MP) contamination is ubiquitous in the environment and many species worldwide have been shown to contain MP. The ecological impact of MP pollution is still unknown, thus there is an urgent need for more knowledge. One key task is to identify species suitable as sentinels for monitoring in key eco-compartments, such as coastal waters. In Norway, mussels (Mytilus spp.) have been monitored for hazardous contaminants through OSPAR since 1981. Norway has the longest coastline in Europe and adding MP to the Norwegian Mussel Watch is therefore important in a European and global context. The present study reports MP data in mussels (332 specimens) collected from multiple sites (n = 15) spanning the whole Norwegian coastline. MPs were detected at all locations, except at one site on the west coast. Among the most surprising findings, mussels from the Barents Sea coastline in the Finnmark region, contained significantly more MPs than mussels from most of the southern part of the country, despite the latter sites being located much closer to major urban areas. Only mussels from a site located very close to Oslo, the capital, contained levels similar to those observed in the remote site in Finnmark. In total an average of 1.5 (±2.3) particles ind-1 and 0.97 (±2.61) particles w.w. g-1 was found. The most common MPs were <1 mm in size, and fibres accounted for 83% of particles identified, although there was inter-site variability. Thirteen different polymeric groups were identified; cellulosic being the most common and black rubbery particles being the second. This study suggests Mytilus spp. are suitable for semi-quantitative and qualitatively monitoring of MPs in coastal waters. However, some uncertainties remain including mussel size as a confounding factor that may influence ingestion, the role of depuration and other fate related processes, and this call for further research.

Comparison of caged and native blue mussels (Mytilus edulis spp.) for environmental monitoring of PAH, PCB and trace metals.

Contaminant bioaccumulation was studied in blue mussels (Mytilus edulis spp.) using the harbor waters of Kristiansand (Norway) as a case study. A suite of chemical contaminants (trace metals, PAHs and PCBs) was analyzed in caged and native mussels as well as in passive samplers (Diffusive Gradients in Thin films (DGT)-devices and silicone rubbers) placed alongside the mussels for estimation of contaminant concentrations in water and uptake rates and bioaccumulation factors (BAFs) in mussels during a six-months deployment period. Estimated logBAFs were in the ranges 2.3-5.5, 3.8-5.2 and 3.2-4.4 for metals, PCBs and PAHs, respectively. Contaminant levels in caged mussels increased rapidly to stable levels for trace metals, whereas for hydrophobic organic contaminants the increase was steady but slow and for many compounds did not reach the levels observed in native mussels. Some key issues related to mussel caging design, such as mussel deployment time and confounding influence from seasonal fluctuations, are discussed herein.

Blue mussels (Mytilus edulis spp.) as sentinel organisms in coastal pollution monitoring: A review.

The blue mussel (Mytilus spp.) is widely used as a bioindicator for monitoring of coastal water pollution (mussel watch programs). Herein we provide a review of this study field with emphasis on: the suitability of Mytilus spp. as environmental sentinels; uptake and bioaccumulation patterns of key pollutant classes; the use of Mytilus spp. in mussel watch programs; recent trends in Norwegian mussel monitoring; environmental quality standards and background concentrations of key contaminants; pollutant effect biomarkers; confounding factors; particulate contaminants (microplastics, engineered nanomaterials); climate change; harmonization of monitoring procedures; and the use of deployed mussels (transplant caging) in pollution monitoring. Lastly, the overall state of the art of blue mussel pollution monitoring is discussed and some important issues for future research and development are highlighted.

May organic pollutants affect fish populations in the North Sea?

The North Sea is a highly productive area with large fish populations that have been extensively harvested over the past century. North Sea fisheries remain important to the surrounding countries despite declining fish stocks over the past decades. The main reason for declining fish stocks is nearly certainly overfishing, but other environmental pressures also affect fish populations, such as eutrophication, climate change, and exposure to metals and organic pollutants, including polyaromatic hydrocarbons (PAHs), alkylphenols, and organochlorine compounds. There are three main sources of organic pollutants in the North Sea: atmospheric, land-based sources, and inputs from offshore gas and oil installations. All three sources contribute to elevated concentrations of organic pollutants in the North Sea compared to the Norwegian Sea. There is evidence that chlorinated organic contaminants were present in sufficiently high concentrations in the southern North Sea two decades ago, to alter embryonal development in fish. The results from extensive, long-term monitoring programs show that some diseases decreased whereas other increased in the southern North Sea and that, among other factors, contaminants may play a role in the temporal changes recorded in disease prevalence. Recent studies demonstrated that components in offshore effluents may affect fish reproduction and that tissues of fish near oil rigs are structurally different to tissues of fish from reference areas. Data on effluents from offshore activities have recently become available through an international workshop (BECPELAG) and follow-up studies.

Environmental effects of the Deepwater Horizon oil spill: A review.

The Deepwater Horizon oil spill constituted an ecosystem-level injury in the northern Gulf of Mexico. Much oil spread at 1100-1300m depth, contaminating and affecting deepwater habitats. Factors such as oil-biodegradation, ocean currents and response measures (dispersants, burning) reduced coastal oiling. Still, >2100km of shoreline and many coastal habitats were affected. Research demonstrates that oiling caused a wide range of biological effects, although worst-case impact scenarios did not materialize. Biomarkers in individual organisms were more informative about oiling stress than population and community indices. Salt marshes and seabird populations were hard hit, but were also quite resilient to oiling effects. Monitoring demonstrated little contamination of seafood. Certain impacts are still understudied, such as effects on seagrass communities. Concerns of long-term impacts remain for large fish species, deep-sea corals, sea turtles and cetaceans. These species and their habitats should continue to receive attention (monitoring and research) for years to come.

An evaluation of two fluorescence screening methods for the determination of chrysene metabolites in fish bile.

Two screening methods, synchronous fluorescence spectrometry (SFS) and high performance liquid chromatography with fluorescence detection (HPLC-F), have been evaluated for their suitability in determining chrysene metabolites in fish bile. The optimal wavelength pair, excitation/emission 272/374 nm, for SFS measurements of chrysene metabolites was identified by analysis of bile taken from fish exposed to the pure compound. This analysis revealed in addition some information about the metabolite pattern. However, when bile from fish exposed to complex and environmentally relevant mixtures of polycyclic aromatic hydrocarbons (PAHs) was analysed using these methods, identification of the chrysene metabolites was poor. Analysis of bile taken from fish exposed to single PAHs identified other three- and four-ring aromatics as the main interfering compounds. Both methods were equally able to discriminate between impacted and reference sites by determination of relative concentrations of fluorescent three- and four-ring aromatics.

Fish bioaccumulation and biomarkers in environmental risk assessment: a review.

In this review, a wide array of bioaccumulation markers and biomarkers, used to demonstrate exposure to and effects of environmental contaminants, has been discussed in relation to their feasibility in environmental risk assessment (ERA). Fish bioaccumulation markers may be applied in order to elucidate the aquatic behavior of environmental contaminants, as bioconcentrators to identify certain substances with low water levels and to assess exposure of aquatic organisms. Since it is virtually impossible to predict the fate of xenobiotic substances with simple partitioning models, the complexity of bioaccumulation should be considered, including toxicokinetics, metabolism, biota-sediment accumulation factors (BSAFs), organ-specific bioaccumulation and bound residues. Since it remains hard to accurately predict bioaccumulation in fish, even with highly sophisticated models, analyses of tissue levels are required. The most promising fish bioaccumulation markers are body burdens of persistent organic pollutants, like PCBs and DDTs. Since PCDD and PCDF levels in fish tissues are very low as compared with the sediment levels, their value as bioaccumulation markers remains questionable. Easily biodegradable compounds, such as PAHs and chlorinated phenols, do not tend to accumulate in fish tissues in quantities that reflect the exposure. Semipermeable membrane devices (SPMDs) have been successfully used to mimic bioaccumulation of hydrophobic organic substances in aquatic organisms. In order to assess exposure to or effects of environmental pollutants on aquatic ecosystems, the following suite of fish biomarkers may be examined: biotransformation enzymes (phase I and II), oxidative stress parameters, biotransformation products, stress proteins, metallothioneins (MTs), MXR proteins, hematological parameters, immunological parameters, reproductive and endocrine parameters, genotoxic parameters, neuromuscular parameters, physiological, histological and morphological parameters. All fish biomarkers are evaluated for their potential use in ERA programs, based upon six criteria that have been proposed in the present paper. This evaluation demonstrates that phase I enzymes (e.g. hepatic EROD and CYP1A), biotransformation products (e.g. biliary PAH metabolites), reproductive parameters (e.g. plasma VTG) and genotoxic parameters (e.g. hepatic DNA adducts) are currently the most valuable fish biomarkers for ERA. The use of biomonitoring methods in the control strategies for chemical pollution has several advantages over chemical monitoring. Many of the biological measurements form the only way of integrating effects on a large number of individual and interactive processes in aquatic organisms. Moreover, biological and biochemical effects may link the bioavailability of the compounds of interest with their concentration at target organs and intrinsic toxicity. The limitations of biomonitoring, such as confounding factors that are not related to pollution, should be carefully considered when interpreting biomarker data. Based upon this overview there is little doubt that measurements of bioaccumulation and biomarker responses in fish from contaminated sites offer great promises for providing information that can contribute to environmental monitoring programs designed for various aspects of ERA.

Environmental risk assessment of combined effects in aquatic ecotoxicology: a discussion paper.

Environmental regulatory edicts within the EU, such as the regulatory framework for chemicals REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals), the Water Framework Directive (WFD), and the Marine Strategy Framework Directive (MSFD) focus mainly on toxicity assessment of individual chemicals although the effect of contaminant mixtures is a matter of increasing concern. This discussion paper provides an overview of the field of combined effects in aquatic ecotoxicology and addresses some of the major challenges related to assessment of combined effects in connection with environmental risk assessment (ERA) and regulation. Potentials and obstacles related to different experimental, modelling and predictive ERA approaches are described. On-going ERA guideline and manual developments in Europe aiming to incorporate combined effects of contaminants, the use of different experimental approaches for providing combined effect data, the involvement of biomarkers to characterize Mode of Action and toxicity pathways and efforts to identify relevant risk scenarios related to combined effects are discussed.

Environmental harm assessment of a wastewater discharge from Hammerfest LNG: a study with biomarkers in mussels (Mytilus sp.) and Atlantic cod (Gadus morhua).

Biologically treated wastewater (WW) from the Hammerfest LNG (liquefied natural gas) plant is discharged to the sea. A study using biomarkers in mussels and Atlantic cod was performed to examine whether this discharge meets a zero harmful emission requirement. Caging of mussels close to the outfall and exposure of mussels and fish to WW in the laboratory were conducted, and a suite of contaminant responsive markers was assessed in exposed animals. In mussels the markers included chemical contaminant levels, haemocyte lysosomal instability and nucleus integrity, cellular energy allocation, digestive gland and gonad histopathology and shell-opening behaviour. In fish, biliary PAH metabolites and gill histopathology biomarkers were measured. A consistent cause-effect relationship between WW treatments and markers measured in test animals was not found. The results therefore indicate that the WW emission is unlikely to represent a significant stress factor for the local marine environment under the conditions studied.