Ageing influences the toxicity of two innovative nanofertilizers to the soil invertebrates Enchytraeus crypticus and Folsomia candida.

The increasing global food demand is threatening the sustainability of agrifood production systems. The intensification of agricultural practices, with inadequate use of pesticides and fertilizers, poses major challenges to the good functioning of agroecosystems and drastically degrades the soil quality. Nanotechnology is expected to optimize the current farming practices and mitigate some associated impacts. Layered double hydroxides (LDHs) are a class of nanomaterials with high potential for use in agricultural productions, mostly due to their sustained release of nutrients. Considering its novelty and lack of studies on the terrestrial ecosystem, it is essential to assess potential long-term harmful consequences to non-target organisms. Our study aimed to evaluate the effect of Zn-Al-NO3 LDH and Mg-Al-NO3 LDH ageing on the survival and reproduction of two soil invertebrate species Enchytraeus crypticus and Folsomia candida. We postulated that the toxicity of nanomaterials to soil invertebrates would change with time, such that the ageing of soil amendments would mediate their impacts on both species. Our results showed that the toxicity of LDHs was species-dependent, with Zn-Al-NO3 LDH being more toxic to E. crypticus, while Mg-Al-NO3 LDH affected more F. candida, especially in the last ageing period, where reproduction was the most sensitive biological parameter. The toxicity of both nanomaterials increased with ageing time, as shown by the decrease of the EC50 values over time. The influence of LDH dissolution and availability of Zn and Mg in the soil pore water was the main factor related to the toxicity, although we cannot rule out the influence of other structural constituents of LDHs (e.g., nitrates and aluminium). This study supports the importance of incorporating ageing in the ecotoxicity testing of nanomaterials, considering their slow release, as effects on soil organisms can change and lead to more severe impacts on the ecosystem functioning.

Soil habitat function after innovative nanoagriproducts application: Effect of ageing on the avoidance behaviour of the soil invertebrates Enchytraeus crypticus and Folsomia candida.

Research on nanotechnology with applications in agriculture has been gathering attention because it may achieve a good balance between agricultural production and environmental integrity. Among the vast nanomaterials, layered double hydroxides (LDHs) are a promising solution for supplying crops with macro- and/or micronutrients. Still, little is known about their safety implications for non-target organisms, such as soil invertebrates. The habitat function of soils might be impacted by potential stressors, which can be assessed through avoidance behaviour tests. This study aimed to assess the effect of two innovative agriproducts, Zn-Al-NO3 LDH and Mg-Al-NO3 LDH, on the avoidance behaviour of the enchytraeid Enchytraeus crypticus and the collembolan Folsomia candida, over time. Simultaneously, Zn and Mg potential release from LDHs to soil was evaluated. Overall, the behaviour of soil invertebrates differed between species, with enchytraeids being more sensitive to LDHs-treated soils than collembolans, possibly explained by their different physiological traits. The behaviour of soil organisms also depended on the LDH structural composition and was time-variable. Soil treated with Zn-Al-NO3 LDH was perceived as less favourable compared to Mg-Al-NO3 LDH, which was preferred to clean soil at most tested concentrations. LDHs toxicity was partly, but not exclusively, related to Zn and Mg release. Cations release over time was demonstrated in the chemical assessment. Still, LDHs toxicity to soil invertebrates decreased as increasing AC50 values were derived over time. Slower dissolution over time might explain the decrease in toxicity. Our study demonstrates that both soil invertebrates could sense LDHs in soil and eventually adapt their behaviour by avoiding or preferring, according to the type and level of LDH present.

Effects of single and combined exposures of gold (nano versus ionic form) and gemfibrozil in a liver organ culture of Sparus aurata.

In vitro methods have gained rising importance in ecotoxicology due to ethical concerns. The aim of this study was to assess the single and combined in vitro effects of gold, as nanoparticle (AuNPs) and ionic (Au+) form, and the pharmaceutical gemfibrozil (GEM). Sparus aurata liver organ culture was exposed to gold (4 to 7200 µg·L-1), GEM (1.5 to 15,000 µg·L-1) and combination 80 µg·L-1 gold +150 µg·L-1 GEM for 24 h. Endpoints related with antioxidant status, peroxidative/genetic damage were assessed. AuNPs caused more effects than Au+, increasing catalase and glutathione reductase activities and damaging DNA and cellular membranes. Effects were dependent on AuNPs size, coating and concentration. GEM damaged DNA at an environmentally relevant concentration, 1.5 µg·L-1. Overall, the effects of the combined exposures were higher than the predicted, based on single exposures. This study showed that liver culture can be a useful model to study contaminants effects.

Impact of Ag2S NPs on soil bacterial community - A terrestrial mesocosm approach.

Soils might be a final sink for Ag2S nanoparticles (NPs). Still, there are limited data on their effects on soil bacterial communities (SBC). To bridge this gap, we investigated the effects of Ag2S NPs (10 mg kg-1 soil) on the structure and function of SBC in a terrestrial indoor mesocosm, using a multi-species design. During 28 days of exposure, the SBC function-related parameters were analysed in terms of enzymatic activity, community level physiological profile, culture of functional bacterial groups [phosphorous-solubilizing bacteria (P-SB) and heterotrophic bacteria (HB)], and SBC structure was analysed by 16S rRNA gene-targeted denaturing gradient gel electrophoresis. The SBC exposed to Ag2S NPs showed a significative decrease of functional parameters, such as ß-glucosidase activity and L-arginine consumption, and increase of the acid phosphatase activity. At the structural level, significantly lower richness and diversity were detected, but at later exposure times compared to the AgNO3 treatment, likely because of a low dissolution rate of Ag2S NPs. In fact, stronger effects were observed in soils spiked with AgNO3, in both functional and structural parameters. Changes in SBC structure seem to negatively correlate with parameters related to phosphorous (acid phosphatase activity) and carbon cycling (abundance of HB, P-SB, and ß-glucosidase activity). Our results indicate a significant effect of Ag2S NPs on SBC, specifically on parameters related to carbon and phosphorous cycling, at doses as low as 10 mg kg-1 soil. These effects were only observed after 28 days, highlighting the importance of long-term exposure experiments for slowly dissolving NPs.

Combined cytotoxic and genotoxic effects of ochratoxin A and fumonisin B1 in human kidney and liver cell models.

Food products can be contaminated by several fungi species and each species may produce different mycotoxins, leading to human combined exposure. Although predictions about the joint toxic effects of mycotoxins can be made from their individual toxicities, experimental data is still limited to allow a reliable hazard assessment. Thus, this study aimed to characterize the combined cytotoxic and genotoxic effects of ochratoxin A (OTA) and fumonisin B1 (FB1) in cell lines representative of their target organs, kidney and liver. Interactions were ascertained using mathematical extensions to the reference models of concentration addition and independent action. Cytotoxicity (MTT assay) data modeling revealed a synergistic pattern for low doses of both FB1 and OTA shifting to antagonism at higher concentration levels, irrespectively of the reference model applied. Concerning genotoxicity assessment, neither OTA nor FB1, individually or in combination, induced a prominent increase in DNA damage (comet assay) or oxidative DNA damage (FPG-comet assay). In conclusion, this study revealed a synergistic cytotoxic effect for OTA and FB1 at low concentration levels. Given that human co-exposure to these two mycotoxins is probable to occur at low doses, these results raise concerns regarding their potential health outcomes that seem to differ from those predicted by an additive model.

Environmental behaviour and ecotoxicity of cationic surfactants towards marine organisms.

Cationic surfactants are surface-active compounds that can be found in many products, including household and cleaning agents. As a consequence, they tend to be discarded into water streams, ultimately ending up in the aquatic environment. In spite of this environmental issue, studies describing their effects towards marine species are lacking. The aim of this study was therefore to evaluate the short-term exposure effects of two commercial cationic surfactants and three novel gemini surfactants on four marine species, the green microalgae Nannochloropsis gaditana and Tetraselmis chuii, the diatom Phaeodactylum tricornutum, and the crustacean Artemia salina. Furthermore, biodegradation and size distribution of the cationic surfactants in artificial seawater were also studied by UV-vis spectrophotometry and dynamic light scattering, respectively. Ecotoxicity tests revealed that the commercial cationic surfactant N-cetyl-N,N,N-trimethylammonium bromide is toxic to all tested marine species while N-dodecyl-N,N,N-trimethylammonium chloride and 1,4-bis-[N-(1-dodecyl)-N,N-dimethylammoniummethyl]benzene dibromide showed the lowest toxicity among the tested cationic surfactants. Besides the novel insights regarding the effects caused by these five cationic surfactants, this work opens prospects for the replacement of commercially available surfactants by more environmentally friendly alternatives.

Effects of gold nanoparticles in gilthead seabream-A proteomic approach.

Despite the widespread use of nanoparticles (NPs), there are still major gaps of knowledge regarding the impact of nanomaterials in the environment and aquatic animals. The present work aimed to study the effects of 7 and 40 nm gold nanoparticles (AuNPs) - citrate and polyvinylpyrrolidone (PVP) coated - on the liver proteome of the estuarine/marine fish gilthead seabream (Sparus aurata). After 96 h, exposure to AuNP elicited alterations on the abundance of 26 proteins, when compared to the control group. AuNPs differentially affected several metabolic pathways in S. aurata liver cells. Among the affected proteins were those related to cytoskeleton and cell structure, gluconeogenesis, amino acids metabolism and several processes related to protein activity (protein synthesis, catabolism, folding and transport). The increased abundance of proteins associated with energy metabolism (ATP synthase subunit beta), stress response (94 kDa glucose-regulated protein) and cytoskeleton structure (actins and tubulins) may represent the first signs of cellular oxidative stress induced by AuNPs. Although higher gold accumulation was found in the liver of S. aurata exposed to 7 nm PVP-AuNPs, the 7 nm cAuNPs were more bioactive, inducing more effects in liver proteome. Gold accumulated more in the spleen than in the other assessed tissues of S. aurata exposed to AuNPs, highlighting its potential role on the elimination of these NPs.

Biological effects and bioaccumulation of gold in gilthead seabream (Sparus aurata) - Nano versus ionic form.

The question of whether gold (Au) is more toxic as nanoparticles or in its ionic form remains unclear and controversial. The present work aimed to clarify the effects of 96 h exposure to 4, 80 and 1600 µg·L-1 of 7 nm gold nanoparticles (AuNPs) - (citrate coated (cAuNPs) or polyvinylpyrrolidone coated (PVP-AuNPs)) - and ionic Au (iAu) on gilthead seabream (Sparus aurata). Effects at different levels of biological organization (behaviour, neurotransmission, biotransformation, oxidative stress/damage and genotoxicity) were assessed. cAuNPs induced oxidative stress and damage (lipid peroxidation increase), even at 4 µg·L-1, and reduced the ability of S. aurata to swim against a water flow at 1600 µg·L-1. Exposure to cAuNPs induced more adverse effects than exposure to PVP-AuNPs. All tested concentrations of Au (nano or ionic form) induced DNA breaks and cytogenetic damage in erythrocytes of S. aurata. Generally, iAu induced significantly more effects in fish than the nano form, probably associated with the significantly higher accumulation in the fish tissues. No fish mortality was observed following exposure to AuNPs, but mortality was observed in the group exposed to 1600 µg·L-1 of iAu.

Multi-generational effects under single and pulse exposure scenarios in two monophyletic Daphnia species.

Anthropogenic activities commonly relate to a set of diffuse and point contamination sources, from industrial, domestic or agricultural outputs, characterized by a chemical cocktail exposure and consequent disturbances of natural ecosystems. Different species may present different sensitivities to contaminants, even when phylogenetically close. This study used two monophyletic Daphnia species from tropical and temperate environments, Daphnia similis and Daphnia magna respectively, to evaluate the variation of their sensitivity to Pb (if any) and fitness during a multi-generational exposure and recovery. To accomplish that, standard acute immobilization tests were done on specific generations. Tests were carried out with exposures to 1) potassium dichromate (K2Cr2O7) to evaluate organisms' sensitivity/fitness, 2) Pb, to monitor variation on Pb sensitivity and 3) the fungicide mancozeb, providing a pulse toxicity approach on generational Pb acclimated daphnids. Since growth is an important trait related to organisms' fitness, organisms' size measurements were also monitored. In addition, organisms were maintained under two different dietary regimes. Our results indicate no variation on daphnids sensitivity to K2Cr2O7, except for D. similis from a recovery period under food restriction. However, a lower Pb sensitivity was seen for both species throughout generations. Both species also showed that under food restriction neonates' sizes were larger than those kept under regular food, while reproduction was considerably reduced. Food restriction also generated opposite outcomes on both species, such as D. magna epigenetic changes and D. similis phenotypic acclimation to Pb. Besides, D. magna pre-exposed to Pb presented lower sensitivity to mancozeb, while the contrary was shown by D. similis. This study indicates that daphnids are capable of acquiring a lower sensitivity to Pb across a long-term exposure, and that Pb pre-exposure can affect the sensitivity to other chemicals. Also, different patterns in multi-generational responses from monophyletic species (especially under oligotrophic media, common on natural habitats) acknowledge the use of representative or native species to assess the effect of contaminants, since monophyletic species can provide different toxicity outputs.

Toxicity of lead and mancozeb differs in two monophyletic Daphnia species.

Lead and mancozeb are two important chemicals used for different human purposes and activities worldwide. Hazard assessment in different areas of the world is carried out with different but phylogenetically similar species, adapted to different climatic conditions, in order to increase relevance. This study evaluated the sensitivity of two monophyletic species, the tropical species Daphnia similis and the temperate species Daphnia magna, to the two chemicals lead and mancozeb. Standard acute and chronic ecotoxicological tests (reproduction and growth), as well as other sublethal measurements such as the intrinsic rate of population increase (r), feeding rate (FR) and O2 consumption, were recorded along with the analysis of the AChE activity to determine the neurotoxicity of both contaminants. Albeit their similar evolutionary status, D. magna generally presented a lower sensitivity to Pb in comparison to D. similis. Despite the differences in sensitivity, both species presented similar patterns of response under Pb exposure, with diminished reproductive outputs, feeding impairment, reduced O2 consumption and no effect on AChE activity. Mancozeb decreased the reproduction, rate of population increase and feeding rate, increased the AChE activity in both species and increased O2 consumption only in D. magna. While D. magna increased O2 consumption under mancozeb exposure, no effects were observed for D. similis. Thus, species may present different responses and sensitivities to different pollutants, regardless of their phylogeny. Therefore, the use of ecotoxicological assays with native species is crucial for a better ecological risk assessment in contaminated areas.

Influence of biochar particle size on biota responses.

Despite the increasing interest for biochar as a soil amendment, a knowledge gap remains on its impacts on non-target soil and aquatic species. We hypothesised that biochar particle size and application rate can play a role in the toxicity to biota. Pine woodchip biochar was incorporated in a clean soil at three particle size classes: small (<0.5 mm), medium (1-2 mm), and large (<4 mm), and at two concentrations: 1% and 6% w/w. A laboratory screening with earthworm Eisenia andrei avoidance behaviour bioassay was carried out to test the most adequate application rates, particle sizes and soil-biochar pre-incubation period. Thereafter, a 28-day greenhouse microcosm experiment was conducted as an ecologically more representative approach. Survival, vertical distribution and weight changes of E. andrei, and bait-lamina consumption were recorded. Soil leachates from the microcosms were collected to evaluate their impact on Daphnia magna immobilisation and Vibrio fischeri (Microtox®) bioluminescence. A feeding experiment with E. andrei was also performed to address earthworm weight changes and to conduct a screening of PAH-type metabolites in their tissue. The 6% <0.5 mm treatment pre-incubated for 96 h induced significant avoidance of the earthworms. Significantly lower bait-lamina consumption was observed in microcosms for the 6% <0.5 mm treatment. Moreover, particle size was a statistically significant factor regarding the loss of weight in the feeding experiment and higher concentration of naphthalene-type metabolites detected in E. andrei tissue, when exposed to <0.5 mm biochar particles. The leachates had no adverse effects on the aquatic species. The results suggest that particles <0.5 mm of pine woodchip biochar can pose sub-lethal effects on soil biota.

Multi-generational exposure to Pb in two monophyletic Daphnia species: Individual, functional and population related endpoints.

To better evaluate chemical damage in chronically contaminated habitats, a nine-generational exposure to Lead (Pb) was done with two monophyletic Daphnia species, from temperate (Daphnia magna) and tropical (Daphnia similis) environments. The multi-generational test consisted generally of a continuous Pb exposed set of organisms, plus an extra control set running simultaneously. To assess daphnids recovery after Pb exposure, some organisms from the sixth generation were transferred to clean media for three extra generations (recovery period; F6 to F9), while others were keep Pb exposed. All setups (control, Pb exposure and recovery period) were submitted to two different dietary regimes, the standard (3 × 105 cells/mL) and restricted food (1.5 × 105 cells/mL) regimes. To evaluate the effects of generational Pb exposure and food regimes, individual, functional and population related endpoints were assessed (number of offspring, body length and rate of population increase (r) and feeding rate (FR)). The tests were conducted on the first (F0) and last generations (F9). No differences were shown on number of offspring and feeding among F9 control and continuous Pb exposed D. magna, although a higher r was shown for F9 Pb exposed organisms. F9 Pb exposed D. similis also presented a higher r than F9 control, however, lethality was induced at high Pb exposure levels. At food restriction the patterns were opposite and D. magna died at high Pb exposure while Pb exposed D. similis was the only setup (compared to control and recovery period) to survive at high Pb exposure levels. Regarding the recovery period, D. magna (standard food) did not cope well with the Pb re-exposure and lethality was induced, while D. similis indicate a decreased Pb sensitivity (only setup that survived high Pb exposure levels). Under food restriction, both species presented a decreased Pb sensitivity and consequent failed recovery (possibly due to epigenetic changes). Both species presented similar patterns regarding generations. Organisms from F0 presented enhanced reproductive outputs in comparison to F9 and the contrary occurred to the FR (even in control organisms). Data show an acclimation under a generational Pb exposure, which could increase the population of adapted organisms in natural habitats. And, since there was not a full recovery after three generations in clean media, an indication of epigenetic changes for both species may also be considered.

Effects and bioaccumulation of gold nanoparticles in the gilthead seabream (Sparus aurata) - Single and combined exposures with gemfibrozil.

Gold nanoparticles (AuNPs) are found in a wide range of applications and therefore expected to present increasing levels in the environment. There is however limited knowledge concerning the potential toxicity of AuNPs as well as their combined effects with other pollutants. Hence, the present study aimed to investigate the effects of AuNPs alone and combined with the pharmaceutical gemfibrozil (GEM) on different biological responses (behaviour, neurotransmission, biotransformation and oxidative stress) in one of the most consumed fish in southern Europe, the seabream Sparus aurata. Fish were exposed for 96 h to waterborne 40 nm AuNPs with two coatings - citrate and polyvinylpyrrolidone (PVP), alone or combined with GEM. Antioxidant defences were induced in liver and gills upon both AuNPs exposure. Decreased swimming performance (1600 µg.L-1) and oxidative damage in gills (4 and 80 µg.L-1) were observed following exposure to polyvinylpyrrolidone coated gold nanoparticles (PVP-AuNPs). Generally, accumulation of gold in fish tissues and deleterious effects in S. aurata were higher for PVP-AuNPs than for cAuNPs exposures. Although AuNPs and GEM combined effects in gills were generally low, in liver, they were higher than the predicted. The accumulation and effects of AuNPs showed to be dependent on the size, coating, surface charge and aggregation/agglomeration state of nanoparticles. Additionally, it was tissue' specific and dependent on the presence of other contaminants. Although, gold intake by humans is expected to not exceed the estimated tolerable daily intake, it is highly recommended to keep it on track due to the increasing use of AuNPs.

Genotoxicity of gold nanoparticles in the gilthead seabream (Sparus aurata) after single exposure and combined with the pharmaceutical gemfibrozil.

Due to their diverse applications, gold nanoparticles (AuNPs) are expected to increase of in the environment, although few studies are available on their mode of action in aquatic organisms. The genotoxicity of AuNPs, alone or combined with the human pharmaceutical gemfibrozil (GEM), an environmental contaminant frequently detected in aquatic systems, including in marine ecosystems, was examined using gilthead seabream erythrocytes as a model system. Fish were exposed for 96 h to 4, 80 and 1600 µg L-1 of 40 nm AuNPs with two coatings - citrate or polyvinylpyrrolidone; GEM (150 µg L-1); and a combination of AuNPs and GEM (80 µg L-1 AuNPs + 150 µg L-1 GEM). AuNPs induced DNA damage and increased nuclear abnormalities levels, with coating showing an important role in the toxicity of AuNPs to fish. The combined exposures of AuNPs and GEM produced an antagonistic response, with observed toxic effects in the mixtures being lower than the predicted. The results raise concern about the safety of AuNPs and demonstrate interactions between them and other contaminants.

Epigenetic variation at the SLC6A4 gene promoter in mother-child pairs with major depressive disorder.

BACKGROUND: Genetic and epigenetic variations of the serotonin transporter gene (SLC6A4) have been related to the etiology of depression. The 5-HTTLPR polymorphism at the SLC6A4 promoter region has two variants, a short allele (S) and a long allele (L), in which the S allele results in lower gene transcription and has been associated with depression. The short S-allele of 5-HTTLPR polymorphism of this gene has been associated with depression. In addition to molecular mechanisms, exposure to early life risk factors such as maternal depression seems to affect the development of depression in postnatal life. The present study investigated the association of 5-HTTLPR polymorphism and CpG DNA methylation (5mC) levels of an AluJb repeat element at the SLC6A4 promoter region in mother-child pairs exposed to maternal depression. METHODS: We analyzed DNA samples from 60 subjects (30 mother-child pairs) split into three groups, with and without major depression disorder (DSM-IV) among children and mothers. The genotyping of 5-HTTLPR polymorphism and quantification of 5mC levels was performed by qualitative PCR and methylation-sensitive restriction enzyme digestion, and real-time quantitative PCR (MSRED-qPCR), respectively. RESULTS: The sample analyzed presented a higher frequency of S allele of 5-HTTLPR (67.5%). Despite the high frequency of this allele, we did not find statistically significant differences between individuals carrying at least one S allele between the depression and healthy control subjects, or among the mother-child pair groups with different patterns of occurrence of depression. In the group where the mother and child were both diagnosed with depression, we found a statistically significant decrease of the 5mC level at the SLC6A4 promoter region. LIMITATIONS: The limitations are the relatively small sample size and lack of gene expression data available for comparison with methylation data. CONCLUSION: In this study, we demonstrated a repeat element specific 5mC level reduction in mother-child pairs, concordant for the diagnosis of depression.

Bioaccumulation and morphological traits in a multi-generation test with two Daphnia species exposed to lead.

Anthropic pressure negatively affects natural environments. Lead (Pb) is a non-essential highly toxic metal that is present in aquatic ecosystems. Two daphnid species from two different latitudes, the temperate Daphnia magna and the tropical Daphnia similis were used as test-organisms to evaluate a long-term Pb exposure. Both species were exposed for nine generations to a chronic concentration of Pb (50 µg/L) and the effects were explored, considering some endpoints not commonly used in toxicity tests: body burden of Pb and presence of granules in the dorsal region of neonates, hemoglobin contents, carapace deformation and morphology, production of males and ephippia (or dormant haploid egg), changes in the eggs' colour and eggs abortion. This multi-generation test was conducted under two food regimes, the usual (3 × 105 cells/mL) and the restricted (1.5 × 105 cells/mL) regime. On generation F6, Pb acclimated neonates were changed to a clean media for three generations, to evaluate exposure retrieval (recovery period). Negative and adverse effects occurred through generations, but no disparity was shown between D. magna and D. similis. The D. magna Pb accumulation showed different patterns regarding food regime. Bioaccumulation was faster under usual food, rapidly reaching a saturation point, whereas a gradual increase occurred under food restriction. A successful retrieval happened regarding Pb in D. magna, since no difference between control and recovering organisms was evidenced regarding their Pb body burdens. Generational Pb exposure led to carapace malformations, Pb aggregation in neonates' dorsal region, reddish extremities, production of males, ephippia (or dormant haploid egg), and aborted eggs, and changes in the eggs' colour (green and white). Food restriction also induced the production of males. Reddish extremities disappeared in recovering organisms and ephippia (or dormant haploid egg) did not occurred during the recovery period. Existent males revealed a shorter lifespan than females (under stress). D. magna and D. similis presented similar responses, for the endpoints analysed; however, it does not mean that this lack of sensitivity difference will be observed when other endpoints (e.g. survival, reproduction) are considered. Bioaccumulation of Pb and adverse effects occurred at the tested concentration of 50µg/L, although higher Pb levels are allowed in the environment as safe concentrations, as reported by the Brazilian legislation and the literature where effects are evidences above 400 µg/L of Pb. Pb effects on reproduction, respiration, malformation, and other adverse effects suggest that a chronic generational exposure can be harmful to both D. magna and D. similis, and that such chronic contaminated environments should not be disregarded when it comes to environmental monitoring.

Correction to: Toxicity prediction and assessment of an environmentally realistic pesticide mixture to Daphnia magna and Raphidocelis subcapitata.

Acknowledgements This work was supported by LEAF (UID/AGR/04129/2013) and CESAM (UID/AMB/50017/2013) from FCT/MEC through national funds, and the cofunding by FEDER (POCI-01-0145-FEDER-007638), within the PT2020 Partnership Agreement and Compete2020.

Hazard assessment of the veterinary pharmaceuticals monensin and nicarbazin using a soil test battery.

Veterinary pharmaceuticals are widely used as food additives in the poultry industry, and the unknown consequences of releasing these compounds into the environment are of concern. The purpose of the present study was to determine the direct impact of 2 veterinary pharmaceuticals (nicarbazin and monensin), commonly used in the poultry industry, on nontarget invertebrates and plant species. Ecotoxicological tests were used to evaluate the acute and chronic toxicity in earthworms (Eisenia andrei), collembolans (Folsomia candida), and 2 plant species (Brassica rapa and Triticum aestivum). Chemical analytical measurements were in good agreement with the nominal concentrations used, although some variability was seen. The results obtained showed no effects of nicarbazin at the highest nominal tested concentration of 1000 mg a.i./kg soil dry weight on any of the organisms, whereas exposure to monensin caused a concentration-specific response pattern. Species sensitivity to monensin decreased in the following rank order: B. rapa > T. aestivum > E. andrei > F. candida, with measured median effect concentrations (based on soil exposure) ranging between approximately 10 and 120 mg/kg. Our results emphasize the importance of using a test battery when assessing ecotoxicological effects by using different ecophysiological endpoints and species from different trophic levels. Environ Toxicol Chem 2018;37:3145-3153. © 2018 SETAC.

A multibiomarker approach highlights effects induced by the human pharmaceutical gemfibrozil to gilthead seabream Sparus aurata.

Lipid regulators are among the most prescribed human pharmaceuticals worldwide. Gemfibrozil, which belongs to this class of pharmaceuticals, is one of the most frequently encountered in the aquatic environment. However, there is limited information concerning the mechanisms involved in gemfibrozil effects to aquatic organisms, particularly to marine organisms. Based on this knowledge gap, the current study aimed to assess biochemical and behavioral effects following a sublethal exposure to gemfibrozil (1.5, 15, 150, 1500 and 15,000 µg L-1) in the estuarine/marine fish Sparus aurata. After the exposure to 1.5 µg L-1 of gemfibrozil, fish had reduced ability to swim against a water flow and increased lipid peroxidation in the liver. At concentrations between 15-15,000 µg L-1, the activities of some enzymes involved in antioxidant defense were induced, appearing to be sufficient to prevent oxidative damage. Depending on the organ, different responses to gemfibrozil were displayed, with enzymes like catalase being more stimulated in gills, whereas glutathione peroxidase was more activated in liver. Although there were no obvious concentration-response relationships, the integrated biomarker response version 2 (IBRv2) analysis revealed that the highest concentrations of gemfibrozil (between 150-15,000 µg L-1) caused more alterations. All the tested concentrations of gemfibrozil induced effects in S. aurata, in terms of behavior and/or oxidative stress responses. Oxidative damage was found at a concentration that is considered environmentally relevant, suggesting a potential of this pharmaceutical to impact fish populations.