Is fish embryo test (FET) according to OECD 236 sensible enough for delivering quality data for effluent risk assessment?

Over the past few years, the fish embryo test (FET) has become widely accepted as an animal-friendly protocol for ecotoxicological research. As Organisation for Economic Co-operation and Development (OECD) 236, the FET has been widely applied for simple mixture exposures under the Registration, Evaluation, Authorisation, and Restriction of Chemicals regulation of the European Union; and now its use is spreading worldwide as a supposedly reliable whole-effluent test (i.e., the testing of complex mixture exposures). However, comparative peer-reviewed data regarding the FET's efficiency for whole-effluent tests are virtually nonexistent. The primary objective of the present study was to make the first comparative test between the FET according to OECD 236 and other standard and slightly modified standard fish protocols used worldwide for whole-effluent tests. For that, we used an untreated hospital effluent considered to be highly toxic but disposed of in municipal sewerage. The base methods were OECD 203 (juvenile), US Environmental Protection Agency method 2000.0 (larvae), and OECD 236 (embryo). We also evaluated the addition of 3 virtually costless sublethal metrics (immobility, nonhatching, and pericardial edema) that could enhance the sensitivity of OECD 236. We observed acute toxicity in all 8 methodologies tested, with a clear escalation in sensitivity (larvae > juvenile ≥ embryo). Larvae were the most sensitive life stage for whole-effluent tests. The addition of sublethal metrics to OECD 236 enhanced its previous sensitivity in over 30%. Thus we conclude that OECD 236 acts below its potential and that the embryonic stage (as used in the FET) may not be the most sensitive life stage for whole-effluent tests. Environ Toxicol Chem 2018;37:2925-2932. © 2018 SETAC.

Assessment of the toxicity of wastewater from the metalworking industry treated using a conventional physico-chemical process.

This article presents results from a toxicity reduction evaluation program intended to describe wastewater from the metalworking industry that was treated using a conventional physico-chemical process. The toxicity of the wastewater for the microcrustacean Daphnia magna was predominantly expressive. Alkaline cyanide wastewater generated from electroplating accounted for the largest number of samples with expressive toxicity. When the raw wastewater concentrations in the batches were repeated, inexpressive toxicity variations were observed more frequently among the coagulated-flocculated samples. At the coagulation-flocculation step, 22.2 % of the treatments had reduced acute toxicity, 30.6 % showed increased toxicity, and 47.2 % remained unchanged. The conductivity and total dissolved solids contents of the wastewater indicated the presence of salts with charges that were inappropriate for the survival of daphnid. The wastewaters treated by neutralization and coagulation-flocculation had average metallic compound contents that were greater than the reference toxic concentrations reported in other studies, suggesting that metals likely contributed to the toxic effects of the wastewater on freshwater microcrustaceans. Thus, alternative coagulants and flocculants should be assessed, and feasible doses should be determined to improve wastewater treatment. In addition, advanced treatment processes should be assessed for their abilities to remove dissolved toxic salts and ions.

Assessment of the freshwater annual fish Cynopoecilus melanotaenia as a toxicity test organism using three reference substances.

This study presents a preliminary evaluation of the use of the Brazilian fish Cynopoecilus melanotaenia as a test organism in toxicity tests. The cryptobiotic stage presented by the eggs of fish C. melanotaenia can overcome the difficulty of continuously keeping cultures and recruiting healthy animals in sufficient numbers to be used in toxicity tests. In order to determine the applicability of this species as a test organism, three different reference substances were evaluated in 96-h acute toxicity tests: Copper sulfate (CuSO4 x 5H2O), sodium dodecil sulfate (C12H25NaO4S), and sodium chloride (NaCl). Sensitivity ranged as follows: copper sulfate (0.05-0.13 mg/L), sodium dodecil sulfate (10.7-19.0 mg/L), and sodium chloride (1.44-1.96 g/L). We conclude that C. melanotaenia shows potential as a test organism in toxicity tests; however, further research should be conducted with other substances and should be compared with the research on other species before we can reach more conclusive results.