Background The Fish Embryo Acute Toxicity (FET) test with the zebrafish (Danio rerio) embryo, the OECD test guideline (TG) 236, has been designed as an alternative for acute fish toxicity testing such as the OECD Acute Fish Toxicity Test (TG 203). To provide equivalent sensitivity to the acute fish test, the original FET test was designed to use only four morphological core endpoints: coagulation of the embryo, lack of somite formation, lack of heart beat, and non-detachment of the tail. These endpoints were selected due to (1) their association with mortality, directly or indirectly, (2) improve the practicality for screening by well-trained technical staff, and (3) the endpoints being relatively simple morphological alterations. Results With the growing need to understand the developmental toxicity of compounds found in the environment, the FET protocol has repeatedly been extended to a multitude of additional morphological endpoints that also allow the monitoring of teratogenicity. As the extensive use of the FET test has generated a multitude of observations in the scientific literature, a harmonisation of the terminology used for the description of the morphological effects seen after chemical exposure has become necessary. Conclusion For this end, the present communication provides an overview of both common and selected more specific morphological effects seen in zebrafish embryos after exposure to a wide variety of chemical substances together with suggestions for a harmonised nomenclature.

Hazard assessment, based on new approach methods (NAM), requires the use of batteries of assays, where individual tests may be contributed by different laboratories. A unified strategy for such collaborative testing is presented. It details all procedures required to allow test information to be usable for integrated hazard assessment, strategic project decisions and/or for regulatory purposes. The EU-ToxRisk project developed a strategy to provide regulatorily valid data, and exemplified this using a panel of > 20 assays (with > 50 individual endpoints), each exposed to 19 well-known test compounds (e.g. rotenone, colchicine, mercury, paracetamol, rifampicine, paraquat, taxol). Examples of strategy implementation are provided for all aspects required to ensure data validity: (i) documentation of test methods in a publicly accessible database; (ii) deposition of standard operating procedures (SOP) at the European Union DB-ALM repository; (iii) test readiness scoring accoding to defined criteria; (iv) disclosure of the pipeline for data processing; (v) link of uncertainty measures and metadata to the data; (vi) definition of test chemicals, their handling and their behavior in test media; (vii) specification of the test purpose and overall evaluation plans. Moreover, data generation was exemplified by providing results from 25 reporter assays. A complete evaluation of the entire test battery will be described elsewhere. A major learning from the retrospective analysis of this large testing project was the need for thorough definitions of the above strategy aspects, ideally in form of a study pre-registration, to allow adequate interpretation of the data and to ensure overall scientific/toxicological validity.

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Filter feeders are target species for microplastic (MP) pollution, as particles can accumulate in the digestive system, disturbing feeding processes and becoming internalized in tissues. MPs may also carry pathogens or pollutants present in the environment. This work assessed the influence of polystyrene (PS) MP size and concentration on accumulation and depuration time and the role of MPs as vectors for metallic (Cd) and organic (benzo(a)pyrene, BaP) pollutants. One-day exposure to pristine MPs induced a concentration-dependent accumulation in the digestive gland (in the stomach and duct lumen), and after 3-day depuration, 45 µm MPs appeared between gill filaments, while 4.5 µm MPs also occurred within gill filaments. After 3-day exposure to contaminated 4.5 µm MPs, mussels showed increased BaP levels whilst Cd accumulation did not occur. Here, PS showed higher affinity to BaP than to Cd. Three-day exposure to pristine or contaminated MPs did not provoke significant alterations in antioxidant and peroxisomal enzyme activities in the gills and digestive gland nor in lysosomal membrane stability. Exposure to dissolved contaminants and to MP-BaP caused histological alterations in the digestive gland. In conclusion, these short-term studies suggest that MPs are ingested and internalized in a size-dependent manner and act as carriers of the persistent organic pollutant BaP.

Environmental context The oil and gas industry has a significant liability in decommissioning offshore infrastructure. Following decommissioning, subsea pipelines could be left on the seabed to provide artificial reefs. Mercury is a contaminant of concern which could remain within pipelines. There are gaps in our knowledge on how mercury moves through the marine environment. We review the current science and identify future research needs to understand potential impacts from mercury in subsea pipelines which will better inform decommissioning activities globally. Abstract In the coming years, the oil and gas industry will have a significant liability in decommissioning offshore infrastructure such as subsea pipelines. The policies around decommissioning vary depending on regional policies and laws. In Australia, the ‘base case’ for decommissioning is removal of all property and the plugging and abandonment of wells in line with the Offshore Petroleum and Greenhouse Gas Storage (OPGGS) Act 2006. Options other than complete removal may be considered where the titleholder can demonstrate that the alternative decommissioning activity delivers equal or better environmental outcomes compared to complete removal and meets all requirements under the OPGGS Act and regulations. Recent research has demonstrated that decommissioning in situ can have significant environmental benefits by forming artificial reefs, increasing marine biodiversity, and providing a potential fishery location. An issue, which has been given less attention, is around contaminants remaining within decommissioned infrastructure and their potential risks to the marine environment. Mercury is a contaminant of concern known to be present in some oil and gas pipelines, but the potential long-term impacts on marine ecosystems are poorly understood. We present a synthesis of information on mercury cycling in the marine environment including key drivers of methylation in sediments and ocean waters, existing models to predict methylmercury concentrations in sediments, and toxicological effects to marine biota. We discuss the applicability of existing water and sediment quality guidelines, and the associated risk assessment frameworks to decommissioning offshore infrastructure contaminated with mercury. Globally, research is needed to provide a comprehensive risk assessment framework for offshore infrastructure decommissioning. We recommend future areas of research to improve our understanding of the potential risks associated with mercury in subsea oil and gas pipelines.

To achieve net zero greenhouse gas emission by 2050 as set out by the 2019 amendment to the 2008 UK Climate Change Act, a major shift towards renewable energy is needed. This includes the development of new methods along with improving and upscaling existing technologies. One example of new methods in

The fish embryo acute toxicity (FET) test with the zebrafish (Danio rerio) embryo according to OECD TG 236 was originally developed as an alternative test method for acute fish toxicity testing according to, e.g., OECD TG 203. Given the versatility of the protocol, however, the FET test has found application beyond acute toxicity testing as a common tool in environmental hazard and risk assessment. Whereas the standard OECD guideline is restricted to four core endpoints (coagulation as well as lack of somite formation, heartbeat, and tail detachment) for simple, rapid assessment of acute toxicity, further endpoints can easily be integrated into the FET test protocol. This has led to the hypothesis that an extended FET test might allow for the identification of different classes of toxicants via a “fingerprint” of morphological observations. To test this hypothesis, the present study investigated a set of 18 compounds with highly diverse modes of action with respect to acute and sublethal endpoints. Especially at higher concentrations, most observations proved toxicant-unspecific. With decreasing concentrations, however, observations declined in number, but gained in specificity. Specific observations may at best be made at test concentrations ≤ EC10. The existence of a “fingerprint” based on morphological observations in the FET is, therefore, highly unlikely in the range of acute toxicity, but cannot be excluded for experiments at sublethal concentrations.