Coelomocytes, immunocompetent cells of lumbricids, have received special attention for ecotoxicological studies due to their sensibility to pollutants. Their in vitro responses are commonly quantified after in vivo exposure to real or spiked soils. Alternatively, quantifications of in vitro responses after in vitro exposure are being studied. Within this framework, the present study aimed at optimizing the neutral red uptake (NRU) assay in primary culture of Eisenia fetida coelomocytes for its application in soil toxicity testing. Optimized assay conditions were: earthworm depuration for 24 h before retrieving coelomocytes by electric extrusion; 2 × 10(5) seeded cells/well (200 µl) for the NRU assay and incubation for 1 h with neutral red dye. Supplementation of the culture medium with serum was not compatible with the NRU assay, but coelomocytes could be maintained with high viability for 3 days in a serum-free medium without replenishment. Thus, primary cultures were used for 24 h in vitro toxicity testing after exposure to different concentrations of Cd, Cu, Ni and Pb (ranging from 0.1 to 100 μg/ml). Primary cultures were sensitive to metals, the viability declining in a dose-dependent manner. The toxicity rank was, from high to low, Pb > Ni > Cd > Cu. Therefore, it can be concluded that the NRU assay in coelomocytes in primary cultures provides a sensitive and prompt response after in vitro exposure to metals.

A B S T R A C TExposure to tricresyl phosphates (TCPs), via for example contaminated cabin air, has been associated with health effects including the so-called aerotoxic syndrome. While TCP neurotoxicity is mainly attributed to ortho-isomers like tri-ortho-cresyl phosphate (ToCP), recent exposure and risk assessments indicate that ToCP levels in cabin air are very low. However, the neurotoxic potential of non-ortho TCP isomers and TCP mixtures is largely unknown. We therefore measured effects of exposure (up to 48 h) to different TCP isomers, mixtures and the metabolite of ToCP (CBDP: cresyl saligenin phosphate) on cell viability and mitochondrial activity, spontaneous neuronal electrical activity, and neurite outgrowth in primary rat cortical neurons.The results demonstrate that exposure to TCPs (24-48 h, up to 10 mM) increases mitochondrial activity, without affecting cell viability. Effects of acute TCP exposure (30 min) on neuronal electrical activity are limited. However, electrical activity is markedly decreased for the majority of TCPs (10 mM) following 48 h exposure. Additional preliminary data indicate that exposure to TCPs (48 h, 10 mM) did not affect the number of neurites per cell or average neurite length, except for TmCP and the analytical TCP mixture (Sigma) that induced a reduction of average neurite length.The combined neurotoxicity data demonstrate that the different TCPs, including ToCP, are roughly equipotent and a clear structure-activity relation is not apparent for the studied endpoints. The noobserved-effect-concentrations (1 mM) are well above current exposure levels indicating limited neurotoxic health risk, although exposures may have been higher in the past. Moreover, prolonged and/or repeated exposure to TCPs may exacerbate the observed neurotoxic effects, which argues for additional research.

The successful treatment of infectious diseases heavily relies on the therapeutic usage of antibiotics. However, the high use of antibiotics in humans and animals leads to increasing pressure on bacterial populations in favour of resistant phenotypes. Antibiotics reach the environment from a variety of emission sources and are being detected at relatively low concentrations. Given the possibility of selective pressure to occur at sub-inhibitory concentrations, the ecological impact of environmental antibiotic levels on microbial communities and resistance levels is vastly unknown. Quantification of antibiotic-resistance genes (ARG) and of antibiotic concentrations is becoming commonplace. Yet, these two parameters are often assessed separately and in a specific spatiotemporal context, thus missing the opportunity to investigate how antibiotics and ARGs relate. Furthermore, antibiotic (multi)resistance has been receiving ever growing attention from researchers, policy-makers, businesses and civil society. Our aim was to collect the limited data on antibiotic concentrations and ARG abundance currently available to explore if a relationship could be defined in surface waters, sediments and wastewaters. A metric of antibiotic selective pressure, i.e. the sum of concentrations corrected for microbial inhibition potency, was used to correlate the presence of antibiotics in the environment to total relative abundance of ARG while controlling for basic sources of non-independent variability, such as country, year, study, sample and antibiotic class. The results of this meta-analysis show a significant statistical effect of antibiotic pressure and type of environmental compartment on the increase of ARG abundance even at very low levels. If global environmental antibiotic pollution continues, ARG abundance is expected to continue as well. Moreover, our analysis emphasizes the importance of integrating existing information particularly when attempting to describe complex relationships with limited mechanistic understanding.

Millions of people rely on active pharmaceutical ingredients (APIs) to prevent and cure a wide variety of illnesses in humans and animals, which has led to a steadily increasing consumption of APIs across the globe and concurrent releases of APIs into the environment. APIs in the environment can have a detrimental impact on wildlife, particularly aquatic wildlife. Therefore, it is essential to assess the potential adverse effects to aquatic ecosystems. The European Water Framework Directive sets out that risk assessment should be performed at the catchment level, crossing borders where needed. The present study defines ecological risk profiles for surface water concentrations of eight APIs (carbamazepine, ciprofloxacin, cyclophosphamide, diclofenac, erythromycin, 17α-ethinylestradiol, metformin and metoprolol) in the Vecht River, a transboundary river that crosses several German and Dutch regions. Ultimately, three main goals were achieved: 1) the geo-referenced estimation of API concentrations in surface water using the GREAT-ER model, 2) the derivation of new predicted no-This article is protected by copyright. All rights reserved. AcceptedArticle effect concentrations (PNECs) for seven of the studied APIs of which three were lower than previously derived values, and 3) the creation of detailed spatially explicit ecological risk profiles of APIs under two distinct water flow scenarios. Under average flow conditions, carbamazepine, diclofenac and 17α-ethinylestradiol were systematically estimated to surpass safe ecological concentration thresholds in at least 68 % of the catchment's water volume. This mounts up to 98% under dry-summer conditions.Graphical abstract. Spatially explicit environmental pharmaceutical risk profiles in the Vecht River basin under two climate condition scenarios.

Active pharmaceutical ingredients (APIs) can reach surface waters used for drinking water extraction and recreational activities, such as swimming and fishing. The aim of the present study was to systematically assess the lifetime human health risks posed by 15 individual APIs and their mixtures occurring in the German-Dutch transboundary Vecht River. An exposure model was developed and used to assess the combined risks of oral and dermal exposure under a variety of exposure conditions. A total of 4500 API uptake values and 165 lifetime risk values were estimated for 15 and 11 APIs, respectively. Overall, the lifetime human health risks posed by the APIs and their mixtures based on modeling results were deemed acceptable under typical exposure conditions. Under very extreme environmental conditions and human behavior, API mixture risks were of potential concern while the risks of individual APIs were negligible, with a few exceptions. The antibiotic doxycycline and analgesic phenazone showed the highest and lowest risks, respectively. The study did not evaluate the potential risks caused by metabolite compounds. Recommendations for water managers are provided to help improve the accuracy and utility of human health risk assessments of pharmaceuticals.