Polluants

Cationic surfactants are used in many industrial processes and in consumer products with concurrent release into the aquatic environment, where they may accumulate in aquatic organisms to regulatoryly relevant thresholds. Here, we aimed to better understand the bioconcentration behavior of three selected cationic surfactants, namely N,N-dimethyldecylamine (T10), N-methyldodecylamine (S12), and N,N,N-trimethyltetradecylammonium cation (Q14), in the cells of fish liver (RTL-W1) and gill (RTgill-W1) cell lines. We conducted full mass balances for bioconcentration tests with the cell cultures, in which the medium, the cell surface, the cells themselves, and the plastic compartment were sampled and quantified for each surfactant by HPLC MS/MS. Accumulation in/to cells correlated with the surfactants' alkyl chain lengths and their membrane lipid–water partitioning coefficient, D_MLW. Cell-derived bioconcentration factors (BCF) of T10 and S12 were within a factor of 3.5 to in vivo BCF obtained from the literature, while the cell-derived BCF values for Q14 were >100 times higher than the in vivo BCF. From our experiments, rainbow trout cell lines appear as a suitable conservative in vitro screening method for bioconcentration assessment of cationic surfactants and are promising for further testing.

Aquatic ecosystems continue to be threatened by chemical pollution. To what extent organisms are able to cope with chemical exposure depends on their ability to display mechanisms of defense across different organs. Among these mechanisms, biotransformation processes represent key physiological responses that facilitate detoxification and reduce the bioaccumulation potential of chemicals. Biotransformation does not only depend on the ability of different organs to display biotransformation enzymes but also on the affinity of chemicals towards these enzymes. In the present study, we explored the ability of different organs and of two freshwater fish to support biotransformation processes through the determination of in vitro phase I and II biotransformation enzyme activity, and their role in supporting intrinsic clearance and the formation of biotransformation products. Three environmentally relevant pollutants were evaluated: the polycyclic aromatic hydrocarbon (PAH) pyrene (as recommended by the OECD 319b test guideline), the fungicide azoxystrobin, and the pharmaceutical propranolol. Comparative studies using S9 sub-cellular fractions derived from the liver, intestine, gills, and brain of brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) revealed significant phase I and II enzyme activity in all organs. However, organ- and species-specific differences were found. In brown trout, significant extrahepatic biotransformation was observed for pyrene but not for azoxystrobin and propranolol. In rainbow trout, the brain appeared to biotransform azoxystrobin. In this same species, propranolol appeared to be biotransformed by the intestine and gills. Biotransformation products could be detected only from hepatic biotransformation, and their profiles and formation rates displayed species-specific patterns and occurred at different magnitudes. Altogether, our findings further contribute to the current understanding of organ-specific biotransformation capacity, beyond the expression and activity of enzymes, and its dependence on specific enzyme-chemical interactions to support mechanisms of defense against exposure.

Diclofenac, a widely used nonsteroidal anti-inflammatory drug (NSAID), enters the aquatic environment worldwide. The effect values available for the derivation of an environmental quality standard (EQS) are markedly heterogeneous, even within the same species. This heterogeneity could partially be attributed to inter-laboratory variation, but is also observed in repeated tests within the same facility. Diclofenac is ionisable; its speciation and potential for uptake and thus toxicity is influenced by pH. A high correlation has previously been observed between effects in zebrafish embryos and the pH-specific partitioning coefficient logD for diclofenac. We hypothesized that the observed heterogeneity could also be attributed to differences in study pH. To test this hypothesis, we reviewed physicochemical data and selected ecotoxicity data that were considered to be reliable and relevant in the latest EU EQS Dossier for which a study pH was reported for further analysis and EQS derivation. We adjusted the reported effect concentrations for differences in uptake using the delta logD value for the worst case pH value of 6.5. pH adjustment of effect values resulted in decreased heterogeneity of the acute effect data and a better fit of the chronic species sensitivity distribution. Both, the MAC-EQS and the AA-EQS were derived using the deterministic approach as data requirements for deriving EQS based on the SSD were not fulfilled. Many studies had to be discarded because test pH was not reported or exposure concentrations had not been analytically verified. Physico-chemical data had to be discarded due to non-relevant experimental conditions or missing information. We strongly encourage scientists publishing ecotoxicity data for ionisable substances to report the test pH together with the effect values and encourage measurement of physico-chemical parameters at environmentally relevant conditions. We recommend to consider adjusting the effect data for ionisable substances according to a worst-case pH in future hazard assessments.