Vitellogenin (Vtg) induction in primary fish hepatocytes has been proposed as an in vitro screening assay for ER agonists and antagonists, but has not yet been used extensively as a high-throughput screening tool due to poor reproducibility, sensitivity and overall feasibility. The present work has evaluated the role of seasonal variation, normalization, optimal culture and assay conditions on the sensitivity, responsiveness and reproducibility of in vitro vtg gene mRNA and protein expression in rainbow trout (Oncorhynchus mykiss) primary hepatocytes using the xenoestrogen 17α-ethynylestradiol (EE2) as a test chemical. The results show that primary hepatocytes display a relatively high individual and seasonal variation in both Vtg mRNA and protein induction potential, although less variance was observed in assay sensitivity. Data normalization of assay response to maximum (3 nM EE2) and minimum (DMSO) Vtg production dramatically reduced this variance and led to improved assay reproducibility. A time-dependent response was observed both for mRNA and protein expression, reaching maximum Vtg induction after 96 h of exposure, although reproducible concentration response curves for both Vtg mRNA and protein could be obtained already after 48 h. A need for chemical re-exposure of the hepatocytes was identified to be important for sustaining exposure concentrations in extended studies (>48 h), whereas different plate formats (96, 24 or 6 wells) did not affect the bioassay performance. In conclusion, standardization of hepatocyte bioassay and test conditions as well as data-normalization procedures are proposed to be instrumental for more consistent and comparable results in future use of this in vitro assay.

Naphthenic acids (NAs) constitute one of the toxic components of the produced water (PW) from offshore oil platforms discharged into the marine environment. We employed liquid chromatography (LC) coupled to high-resolution mass spectrometry with electrospray ionization (ESI) in negative mode for the comprehensive chemical characterization and quantification of NAs in PW samples from six different Norwegian offshore oil platforms. In total, we detected 55 unique NA isomer groups, out of the 181 screened homologous groups, across all tested samples. The frequency of detected NAs in the samples varied between 14 and 44 isomer groups. Principal component analysis (PCA) indicated a clear distinction of the PW from the tested platforms based on the distribution of NAs in these samples. The averaged total concentration of NAs varied between 6 and 56 mg L–1, among the tested platforms, whereas the concentrations of the individual NA isomer groups ranged between 0.2 and 44 mg L–1. Based on both the distribution and the concentration of NAs in the samples, the C8H14O2 isomer group appeared to be a reasonable indicator of the presence and the total concentration of NAs in the samples with a Pearson correlation coefficient of 0.89.

Life-history traits determine individual fitness and the fate of populations. Imidacloprid, a widely used neonicotinoid insecticide, which persists in soil for more than 100 d at biologically relevant levels, may affect nontarget and ecologically important species, such as collembolans. In the present study, we determined the sublethal effects of shortterm imidacloprid exposure and postexposure recovery in the collembolan Folsomia quadrioculata, which occurs abundantly across the northern hemisphere. We assessed survival, egg production, and hatching success in adult springtails exposed for 14 d through the diet to imidacloprid, followed by a 28-d postexposure phase. Survival and hatching success were high throughout the experiment in all the treatments, with no clear concentration dependence. However, egg production declined during the exposure phase and nearly stopped between 8 and 14 d in all the treatments (except the control) but resumed during the postexposure phase. Moreover, the resumption of egg production showed a concentration-dependent delay. Our findings suggest that low imidacloprid exposures can restrict reproduction, with potentially severe consequences for the population, notwithstanding the partial recovery in egg production.

Emamectin benzoate (EMB) is an antisea lice chemical widely used in the aquaculture that may also unintentionally affect nontarget crustaceans in the environment. Although the adverse effects of this compound are well documented in various species, the full modes of action (MoAs) are still not well characterized. The current study was therefore conducted to characterize the MoAs of EMB and link perturbations of key toxicological pathways to adverse effects in the model freshwater crustacean Daphnia magna. Effects on molting and survival were determined after 48 h exposure to EMB, whereas global transcriptional changes and the ecdysone receptor (EcR) binding potency was determined to characterize the MoA. The results showed that the molting frequency and survival of D. magna decreased in a concentration-dependent manner, and the observed changes could not be attributed to direct interactions with the EcR. Major MoAs such as activation of glutamate-gated chloride channels and gamma-aminobutyric acid signaling, disruption of neuroendocrine regulation of molting, perturbation of energy homeostasis, suppression of DNA repair and induction of programmed cell death were observed by transcriptional analysis and successfully linked to the adverse effects. This study has demonstrated that acute exposure to intermediate and high pM levels of EMB may pose hazards to nontarget crustaceans in the aquatic environment.

Collembola (springtails) are important members of soil communities worldwide by contributing to degradation of organic matter. In nature, Collembola might be exposed to the neonicotinoid insecticide imidacloprid, which is fairly persistent in soil. We exposed the widespread Hypogastrura viatica to imidacloprid through soil or food, and monitored the animals during exposure and a post-exposure period. We recorded effects on life-history traits affecting individual fitness; that is, mortality, behavioral activity, several reproduction traits, and moulting frequency.Exposure through soil led to a concentration-dependent mortality, while the mortality from dietary exposure possibly reflected reduced feeding activity. The body burden of This article is protected by copyright. All rights reserved. Accepted Articleimidacloprid in the Collembola did not differ between treatments. We found no sign of recovery in behavioral activity following exposure in neither experiment. The egg production of H. viatica was not significantly affected by imidacloprid at 0.01 mg/kg dry soil, but showed a tendency to reduce number of eggs per batch and reduced hatching success. At higher concentrations, the reproduction was close to, or completely stopped. The moulting frequency decreased during exposure, while in the post-exposure period, we saw milder effects in the highest concentrations, suggesting elimination through moulting or reduced toxic response due to reduced feeding activity. Overall, H. viatica was more sensitive to imidacloprid than previously studied Collembola, which highlights the importance of considering species sensitivities when risk assessing soil environments.