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Soil water repellency (SWR) significantly affects plant growth, along with surface and subsurface hydrology, posing a challenge for agricultural productivity and environmental sustainability. Nowadays, the occurrence of microplastics (MP) in the environment, particularly from agricultural practices, raises concerns about MP impact on soil properties. Among them, SWR is affected by hydrophobicity of MP particles detected in soils. This study introduces a method and presents results of a screening test to assess the effects of MP on SWR, utilizing Water Drop Penetration Time (WDPT) analysis under controlled laboratory conditions in destructed soil samples. We compared SWR of two soil types differing in portion of sand, loam and clay. Soils were mixed with three different types of MP originating from agricultural mulch films: low-density polyethylene (LDPE), biodegradable polybutylene adipate terephthalate (PBAT), and starch-based biodegradable plastics (Starch). The MP were milled to a uniform size range of some 10 to 300 μm and mixed with the soil samples. WDPT measurements were taken immediately after mixing and recorded for up to 60 s in order to find MP concentration levels at which strongly or more severely water repellency is inducted on soil samples. Our findings reveal that both, soil type and MP type significantly influence SWR, where there are notable differences observed between bio-based (Starch based) and non-bio-based (LDPE and PBAT) plastics’ effects on SWR in the two tested soil types. Data highlights the distinct behaviour of Starch in altering soil hydrophobicity, prominently different from the impact of both PBAT and LDPE. The measurement technique we have developed for quantifying SWR levels could be used for both research applications and the dissemination of findings. It can significantly enhance decision-making processes regarding the selection of optimal plastic alternatives for agricultural use.

Agricultural mulching films are potential sources of microplastics (MPs) in soil. As an alternative to conventional non-degradable mulching films, a variety of different biodegradable mulching films are used. However, it is not yet known whether MPs from biodegradable mulching films pose a lower risk to terrestrial invertebrates compared to MPs from conventional mulching films. In this study, the effects of MPs produced from two conventional polyethylene (PE-1 and PE-2) and two biodegradable (starch-based poly(butylene adipate co-terephthalate); PBAT-BD-1, and PBAT-BD-2) fossil-based mulching films on terrestrial crustacean woodlice Porcellio scaber and mealworm Tenebrio molitor were compared. A key finding was that no clear differences in induced responses between biodegradable and conventional MPs were detected. No adverse effects on P. scaber after two weeks and on T. molitor after four weeks of exposure were observed up to 5 % (w/w dry soil) of either MP type. However, some sublethal physiological changes in metabolic rate and immune parameters were found in P. scaber after two weeks of exposure indicating a response of organisms to the presence of MP exposure in soil. In addition, it was demonstrated that both types of MPs might affect the soil water holding capacity and pH. In conclusion, we confirmed that biodegradable MPs can induce responses in organisms hence further studies testing the environmental hazard of biodegradable MPs are justified., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Physico-chemical properties of microplastics (MPs) change during weathering in the environment. There is a lack of knowledge about the effects of such environmentally relevant MPs on organisms. We investigated: 1) the physico-chemical changes of MPs due to UV-VIS weathering, and 2) compared the effect of pristine and aged MPs on the behaviour of the water flea Daphnia magna and terrestrial crustacean Porcellio scaber. Dry powders of MPs were produced from widely used polymer types: disposable three-layer polypropylene (PP) medical masks (inner, middle and outer), polyester textile fibres, car tires and low-density polyethylene (LDPE) bags and were subjected to accelerated ultraviolet-visible (UV-VIS) ageing. Our results show that the extent of transformation depends on the type of polymer, with PP showing the most changes, followed by LDPE, textile fibres and tire particles. Obvious fragmentation was observed in PP and textile fibres. In the case of PP, but not polyester textile fibres, changes in FTIR spectra and surface properties were observed. Tire particles and LDPE did not change in size, but clear changes were observed in their FTIR spectra. Most MPs, aged and pristine, did not affect the swimming of daphnids. The only effect observed was a significant increase in path length and swimming speed for the pristine tire particles when the recording was done with particles remaining in the wells. After transfer to a clean medium, this effect was no longer present, suggesting a physical rather than chemical effect. Similarly, woodlice showed no significant avoidance response to the MPs tested, although there was a noticeable trend to avoid soils contaminated with pristine polyester textile fibers and preference towards the soils contaminated with aged MP of the middle mask layer. Overall, the apparent changes in physico-chemical properties of MPs after accelerated ageing were not reflected in their effects on woodlice and daphnids., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Mulching films, widely used in agriculture, are a large source of microplastics (MPs) to soil. However, there is little knowledge on the long-term effects of agricultural MPs on soil invertebrates. We investigated the effects of MPs from conventional non-biodegradable, fossil-based, low-density polyethylene (PE) and biodegradable fossil-based poly(butylene adipate-coterephthalate) (starch-PBAT blend) mulching films on two generations of the mealworm Tenebrio molitor. No effects of MPs (0.005 %-5 %, w/w dry food) on mealworm development and survival were observed until the end of the experiments (12 weeks for the first generation, nine weeks for the second generation), but effects on their moulting and growth were observed. These were most evident for PE MPs (5 %, w/w), where a decrease in larval growth and moulting was noted in the first generation. On the contrary, PBAT MPs (5 %, w/w) significantly induced the growth of mealworms in the second generation. In addition, there was a non-significant trend towards increased growth at all other PBAT MP exposure concentrations. Increased growth is most likely due to the biodegradation of starch PBAT MPs by mealworms. Overall, these data suggest that PE and PBAT MPs do not induce significant effects on mealworms at environmentally relevant concentrations, but rather only at very high exposure concentrations (5 %)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The exposure of organisms to microplastics could compromise their ability to cope with other environmental stressors, such as infections. In this context, we investigated the effects of a 14-day exposure of the terrestrial isopod Porcellio scaber to tire particles in soil (1.5 % w w -1 dry weight) on the organisms' response to a secondary exposure, i.e., injection of the bacterial endotoxin lipopolysaccharide. In addition, the insecticide chlorpyrifos (2 mg kg -1 dry weight) was tested as a positive control. The survival and immune response of P. scaber was assessed at the end of the 7- and 14-day primary exposure and two days after the secondary exposure, by analyzing selected haemolymph immune parameters (total haemocyte count, differential haemocyte count, and haemocyte viability). No change in survival was observed after primary exposure of P. scaber to tire particles or chlorpyrifos. However, primary exposure to chlorpyrifos triggered a strong activation of the immune response, which was not the case following exposure to the tire particles. Further injection of lipopolysaccharide into the body did not affect the survival of animals exposed to tire particles or chlorpyrifos, while a strong immunomodulatory change was observed, particularly with chlorpyrifos, and to some extent, tire particles. Based on these results, we conclude that exposure of P. scaber to tire particles or chlorpyrifos has no significant effect on the susceptibility of the organism to lipopolysaccharide in terms of their mortality, but primary exposure to an insecticide significantly modulates the immune response of the organisms to a second stressor. We discuss the "stress on stress" approach for testing low-toxic substances, such as microplastics, where an environmentally realistic exposure is followed by a secondary exposure., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

This study investigated impacts of microplastics from disposable polypropylene medical masks on woodlice Porcellio scaber, mealworm larvae Tenebrio molitor and enchytraeids Enchytraeus crypticus. Effects of microplastics on survival, reproduction, immune parameters and energy-related traits were assessed after 21 days exposure in soil. Microplastics obtained from each medical mask layer separately differed in size and shape (inner frontal layer: 45.1 ± 21.5 µm, fibers; middle filtering layer: 55.6 ± 28.5 µm, fragments; outer layer: 42.0 ± 17.8 µm, fibers) and composition of additives. Overall, the concentrations of metals and organic chemicals were too low to cause effects on soil invertebrates. The microplastics from disposable medical masks at 0.06%, 0.5%, 1.5%, w/w did not induce severe adverse effects on survival or reproduction (for enchytraeids). A transient immune response of woodlice and a change in energy-related traits in mealworms were observed, which was most clearly seen for the microplastics from the outer layer. This was reflected in increased electron transfer system activity of mealworms and different immune response dynamics of woodlice. In conclusion, the tested soil invertebrates respond to microplastics from disposable medical masks, but it remains unclear what these changes mean for their fitness on the long term., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The presence of microplastics (MPs) in the environment has raised many concerns, and therefore approaches and technologies to remove them in situ are of high interest. In this context, we investigated the interactions between polyethylene MPs (fragments with a mean size of 149 ± 75 μm) and an aquatic floating macrophyte Lemna minor in order to assess its potential use for in situ phytoremediation. We first investigated the long-term effects of a high (100 mg/L = 9600 MPs/L), but still environmentally relevant concentration of MPs on L. minor. Subsequently bioadhesion of MPs was studied and the number and strength of MPs adhering to plant biomass were assessed. MPs did not adversely affect various parameters of plants (e.g., specific growth rate, chlorophyll contents, total antioxidant capacity, electron transport system activity, and contents of energy-rich molecules) throughout the duration of the experiment (12 weeks), except for the first week of the experiment, when protein content and total antioxidant capacity were affected. On the other hand, MPs affected the root length of L. minor during the first eight weeks of the experiment, while further exposure resulted in a decrease in the effects, indicating the ability of L. minor to tolerate the presence of MPs for a long period of time. MPs adhered rapidly to the plant biomass and the average percentages of strongly and weakly adhered particles were 6.5% and 20.0%, respectively, of the total MPs applied. In summary, results of this study suggest that L. minor can tolerate hotspot concentrations of MPs and can collect MPs from the water surface. Therefore, phytoremediation using floating plants could be considered as a potential method for in situ removal of MPs from the aquatic environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Secondary salinization of freshwater is becoming a growing environmental problem. Currently, there is few data available on the effects of salinisation on subterranean crustaceans that are vital for the maintenance of groundwater ecosystem functioning. In this study, the sensitivity of subterranean Niphargus amphipods to NaCl was investigated. We expected that cave-dwelling species would be more sensitive as surface-subterranean boundary species. Eight ecologically different Niphargus species were tested: four live at the boundary between the surface and subterranean ecosystems (N. timavi, N. krameri, N. sphagnicolus, N. spinulifemur), three live in cave streams (N. stygius, N. scopicauda, N. podpecanus), and one species (N. hebereri) lives in anchialine caves and wells. The organisms were exposed to five concentrations of NaCl for 96 h and afterwards the immobility, mortality, and electron transfer system (ETS) activity (a measure for metabolic rate of animals) were evaluated. As expected, the most tolerant species was N. hebereri dwelling in naturally high-salinity habitat. However, contrary to our expectations, the species collected at the surface-subterranean boundary were more sensitive as cave stream species when their immobility and mortality were assessed. Interestingly, the majority of Niphargus tested were more NaCl tolerant as can be deduced from currently available data for subterranean and surface crustaceans. We could not observe a clear trend in ETS activity changes between groups of surface-subterranean boundary and cave streams species after exposure to NaCl stress, but it appears that osmotic stress-induced metabolic rate changes are species-specific. This study shows that amphipods Niphargus can be a valuable subterranean environmental research model and further ecotoxicity research is of interest., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Microplastics are very common contaminants in the environment. Despite increasing efforts to assess the effects of microplastics on soil organisms, there remains a lack of knowledge on how organisms respond to diverse types of microplastics after different exposure durations. In the present study, we investigated the immune response of the terrestrial crustacean Porcellio scaber exposed to the two most common microplastic particles in the environment: polyester fibres and tyre particles. We also tested two natural particles: wood dust and silica powder, with all treatments performed at 1.5% w/w. The response of P. scaber was evaluated at the level of the immune system, and also the biochemical, organism and population level, after different exposure durations (1, 2, 4, 7, 14, 21 days). These data reveal dynamic changes in the levels of some immune parameters shortly after exposure, with a gradual return to control values. The total number of haemocytes was significantly decreased after 4 days of exposure to tyre particles, while the proportion of different haemocyte types in the haemolymph was altered shortly after exposure to both polyester fibres and tyre particles. Moreover, 7 days of exposure to tyre particles resulted in increased superoxide dismutase activity in the haemolymph, while metabolic activity in whole woodlice (measured as electron transport system activity) was increased after exposure for 7, 14 and 21 days. In contrast, the natural particles did not elicit any significant changes in the measured parameters. Survival and feeding of P. scaber were not altered by exposure to the microplastics and natural particles in soil. Overall, this study defines a time-dependent transient immune response of P. scaber, which indicates that immune parameters represent sensitive biomarkers of exposure to microplastics. We discuss the importance of using natural particles in studies of microplastics exposure and their effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

It is becoming increasingly important to develop assessment criteria for the quality of nanoplastics studies. This study is an attempt to establish such criteria based on those developed for engineered nanomaterials, the GUIDEnano and DaNa criteria being two representatives. These criteria were applied to studies on polystyrene nanoparticles (PS NPs), which currently represent the majority of studies on nanoplastics. We compiled a list of existing nanomaterial-related criteria that are not fully relevant to PS NPs and propose additional nanoplastic-specific criteria targeting polymer chemical composition, source, production and field collection, impurities/chemical additives, density, hydrophobicity, colour, and chemical leaching. For each criterion, scientific justification is provided. We conclude that the existing study quality assessments originally developed for nano(eco)toxicity studies can, through refinements, be applied to those dealing with nanoplastics studies, with a further outlook on microplastics. The final quality criteria catalogue presented here is intended as a starting point for further elaborations considering different purposes of an assessment., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Microplastics and agrochemicals are common pollutants in terrestrial ecosystems. Their interaction during coexistence in soils may influence their fate and adverse effects on terrestrial organisms. The aim of this study was to investigate how the exposure to two types of microplastics; polyester fibres, and crumb rubber; induce changes in immune parameters of Porcellio scaber and if the co-exposure of microplastics affects the response induced by the organophosphate pesticide chlorpyrifos. A number of immune parameters, such as total haemocyte count, differential haemocyte count, and phenoloxidase-like activity were assessed. In addition, the acetylcholinesterase (AChE) activity in the haemolymph was evaluated as a measure of the bioavailability of chlorpyrifos. After three weeks of exposure, the most noticeable changes in the measured immune parameters and also a significantly reduced AChE activity were seen in chlorpyrifos-exposed animals. Both types of microplastic at environmentally relevant concentrations caused only slight changes in immune parameters which were not dependent on the type of microplastic, although the two types differed significantly in terms of the chemical complexity of the additives. Mixtures of chlorpyrifos and microplastics induced changes that differed from individual exposures. For example, alterations in some measured parameters suggested a reduced bioavailability of chlorpyrifos (AChE activity, haemocyte viability) caused by both types of microplastics exposure, but the increase of haemocyte count was promoted by the presence of fibres implying their joint action. In conclusion, this study suggests that immune processes in P. scaber are slightly changed upon exposure to both types of microplastics and microplastics can significantly modulate the effects of other co-exposed chemicals. Further research is needed on the short-term and long-term joint effects of microplastics and agrochemicals on the immunity of soil invertebrates., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Environmental stressors are assessed through methods that quantify their impacts on a wide range of metrics including species density, growth rates, reproduction, behaviour and physiology, as on host-pathogen interactions and immunocompetence. Environmental stress may induce additional sublethal effects, like mutations and epigenetic signatures affecting offspring via germline mediated transgenerational inheritance, shaping phenotypic plasticity, increasing disease susceptibility, tissue pathologies, changes in social behaviour and biological invasions. The growing diversity of pollutants released into aquatic environments requires the development of a reliable, standardised and 3R (replacement, reduction and refinement of animals in research) compliant in vitro toolbox. The tools have to be in line with REACH regulation 1907/2006/EC, aiming to improve strategies for potential ecotoxicological risks assessment and monitoring of chemicals threatening human health and aquatic environments. Aquatic invertebrates' adult stem cells (ASCs) are numerous and can be pluripotent, as illustrated by high regeneration ability documented in many of these taxa. This is of further importance as in many aquatic invertebrate taxa, ASCs are able to differentiate into germ cells. Here we propose that ASCs from key aquatic invertebrates may be harnessed for applicable and standardised new tests in ecotoxicology. As part of this approach, a battery of modern techniques and endpoints are proposed to be tested for their ability to correctly identify environmental stresses posed by emerging contaminants in aquatic environments. Consequently, we briefly describe the current status of the available toxicity testing and biota-based monitoring strategies in aquatic environmental ecotoxicology and highlight some of the associated open issues such as replicability, consistency and reliability in the outcomes, for understanding and assessing the impacts of various chemicals on organisms and on the entire aquatic environment. Following this, we describe the benefits of aquatic invertebrate ASC-based tools for better addressing ecotoxicological questions, along with the current obstacles and possible overhaul approaches., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Studying the behaviour of nanomaterials after their release into natural water is essential to understand the risk associated to their environmental exposure. In particular, the interaction and adsorption of dissolved organic matter onto nanoparticles strongly influence the behaviour and fate of nanomaterials in natural water systems. We herein study the interaction of Au and Ag nanoparticles and humic acids, the principal component of natural dissolved organic matter. Physicochemical characterization results showed the formation of an organic matter corona, consisting of two layers: a "hard" one, firmly bound to the nanoparticle surface, and a "soft" one, in dynamic equilibrium and, consequently, highly dependent on the media organic matter concentration. The extent of the electro-steric stabilization of the so called environmental corona depends on the size of the supramolecular association of humic acid (which depends on its hydrophilic and lipophilic moieties), the nanoparticle size, the total concentration of organic matter in the media, and the ratio between them. Interestingly, environmental coronas can eventually prevent Ca 2+ and Mg 2+ induced aggregation at concentrations range present in most of the freshwater bodies. The humic coating formed on top of the Au or control Ag nanoparticles presented a similar profile, but the corrodibility of Ag led to a more natural detachment of the corona. These results were further confirmed by exposing the nanoparticles to a model of natural water and standard mud (LUFA 2.2 dispersion). In the latter case, after several days, nanoparticle sedimentation was observed, which was attributed to interactions with macro organic and inorganic matter (fraction larger than particulate matter)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

One of the most widely used aquatic standarized tests for the toxicity screening of chemicals is the acute toxicity test with the freshwater crustacean Daphnia magna, which has also been applied in the toxicity screening of manufactured nanoparticles (NPs). However, in the case of non-soluble NPs most of the results of this test have showed no effect. The aim of the work presented here was to modify the standardized test by the least possible extent to make it more sensitive for non-soluble particles. The standard acute immobilisation assay with daphnids was modified by prolonging the exposure period and by measuring additional endpoints. Daphnids were exposed to TiO 2 NPs in a standard acute test (48h of exposure), a standard acute test (48h of exposure) followed by 24h recovery period in clean medium or a prolonged exposure in the NPs solutions totaling 72h. Together with immobility, the adsorption of NPs to body surfaces was also observed as an alternative measure of the NPs effects. Our results showed almost no effect of TiO 2 NPs on D. magna after the 48h standard acute test, while immobility was increased when the exposure period to TiO 2 NPs was prolonged from 48h to 72h. Even when daphnids were transferred to clean medium for additional 24h after 48h of exposure to TiO 2 NPs the immobility increased. We conclude that by transferring the daphnids to clean medium at the end of the 48h exposure to TiO 2 NPs, the delayed effects of the tested material can be seen. This methodological step could improve the sensitivity of D. magna test as a model in nanomaterial environmental risk assessment., (Copyright © 2018 Elsevier Inc. All rights reserved.)

We investigated the effects of a commercial pigment grade rutile TiO2 on the terrestrial isopod Porcellio scaber in three locations that differed in terms of abiotic and biotic conditions: the laboratory, open air, and the closed barn. Mortality and isopod energy reserves (digestive gland total proteins, lipids and carbohydrates) were not affected following 14days exposure to up to 1000mg TiO2 per kg dry leaves (mg/kg) under any experimental scenario. However, in the field tests, isopods consumption of TiO2-coated leaves was reduced compared to that of uncoated leaves and the decrease was not dose-dependent. The highest reduction was in the closed barn (45-56%) rather than in the open-air (38-40%). In laboratory-based food choice tests, isopods neither preferred nor avoided leaves coated with TiO2, suggesting that rather than sensing the TiO2 on the leaves directly, the isopods under open-air and barn exposure responded to altered attractiveness and/or palatability of the TiO2 amended leaves. We propose that this could be due to altered microbial population on the leaves, a hypothesis that requires further investigation. Although short-term exposure to atmospheric deposition of up to 1000mg/kg commercial TiO2 is unlikely to pose an immediate threat to isopod mortality and energy balance, reduced leaf feeding may have implications for the decomposition of plant material., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Sterilization is the final surface treatment procedure of all implantable devices and is one of the key factors which have to be considered before implementation. Since different sterilization procedures for all implantable devices influence mechanical properties as well as biological response, the influence of different sterilization techniques on titanium nanotubes was studied. Commonly used sterilization techniques such as autoclaving, ultra-violet light sterilization, hydrogen peroxide plasma sterilization as well as the not so frequently used gaseous oxygen plasma sterilization were used. Three different nanotube diameters; 15 nm, 50 nm and 100 nm were employed to study the effects of various sterilization techniques. It was observed that autoclave sterilization resulted in destruction of nanotubular features on all three studied nanotube diameters, while UV-light and both kinds of plasma sterilization did not cause any significant morphological changes on the surfaces. Differences between the sterilization techniques employed influenced cytocompatibility, especially in the case of nanotubes with 100 nm diameter., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In arthropods, the determination of two important parameters of digestive juices, i.e. the total surfactant concentration and the critical micelle concentration (CMC), is challenging due to small sample volumes and low surfactant concentrations. In this work, we report a successful implementation of potentiometric titrations using the surfactant ion-selective electrode (SISE) and the pyrene fluorescence method (PFM) for the determination of the total surfactant concentration and CMC in the digestive juice of terrestrial isopod crustaceans Porcellio scaber. Pooled digestive juice extracts of four (SISE) or two (PFM) animals were used per measurement run. In both cases, digestive juice extracts in 100 μL of deionized water were sufficient for one measurement run. The total surfactant concentration of P. scaber digestive juice was determined to be 9.2 ± 3.5mM and the CMC was approximately 90 μM. Our work presents an important improvement towards easy CMC determination in small volume samples in comparison with the commonly used stalagmometric technique, where much larger sample volumes are usually needed. To date, the total surfactant concentration was not measured in the digestive juices of arthropods other than Homarus vulgaris, Astacus leptodactylus and Cancer pagurus, for which complex separation and analytical techniques were required. Our results obtained by SISE and PFM therefore present the first successful quantification of surfactants and their CMC in small volumes of arthropod digestive juice without prior separation or purification techniques., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The aim of this study was to determine whether citric acid adsorbed onto cobalt ferrite (CoFe2O4) nanoparticles (NPs) influences the bioavailability of their constituents Co and Fe. Dissolution of Co and Fe was assessed by two measures: (i) in aqueous suspension using chemical analysis, prior to application onto the food of test organisms; and (ii) in vivo, measuring the bioavailability in the model terrestrial invertebrate (Porcellio scaber, Isopoda, Crustacea). The isopods were exposed to citric-acid-adsorbed CoFe2O4 NPs for 2 weeks, and tissue accumulation of Co and Fe was assessed. This was compared to pristine CoFe2O4 NPs, and CoCl2 and Fe(III) salts as positive controls. The combined data shows that citric acid enhances free metal ion concentration from CoFe2O4 NPs in aqueous suspension, although in vivo, very similar amounts of assimilated Co were found in isopods exposed to both types of NPs. Therefore, evaluation of the dissolution in suspension by chemical means is not a good predictor of metal assimilation of this model organism; body assimilation of Co and Fe is rather governed by the physiological capacity of P. scaber for the uptake of these metals. Moreover, we propose that citric acid, due to its chelating properties, may hinder the uptake of Co that dissolves from citric-acid-adsorbed CoFe2O4 NPs, if citric acid is present in sufficient quantity., (Copyright © 2014 Elsevier B.V. All rights reserved.)

We provide data obtained by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) on the interaction of ingested tungsten nanofibers with epithelial cells of the digestive tubes of a test organism Porcellio scaber. Conventional toxicity endpoints including feeding behaviour, weight loss and mortality were also measured in each investigated animal. No toxicity was detected in any of exposed animals after 14 days of feeding on tungsten nanofiber dosed food, but when nanofibers enter the digestive system they can react with epithelial cells of the digestive tubes, becoming physically inserted into the cells. In this way, nanofibers can injure the epithelial cells of digestive gland tubes when they are ingested with food. Our SEM data suggest that peristaltic forces may have an important role, not predicted by in vitro experiments, in the interactions of nanomaterials with digestive intestinal cells., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The aim of this work was to investigate if the activities of catalase and glutathione S-transferase in a control population of terrestrial isopods (Porcellio scaber) are correlated with the physiological condition of the isopods. For this purpose, the activities of these enzymes were analysed in isopods from a stock population and in parallel, the physiological condition of the same specimens was assessed using a histological approach based on epithelial thickness and lipid droplets. We found a correlation between antioxidant enzymes and the physiological condition of the isopods. This implies that these enzymes could be used as predictive indicators of the physiological condition in a stock population before comprehensive toxicological studies are conducted and also in control group after the experiment. When a control group is found to be very heterogeneous in terms of physiological condition, the experiment should be repeated with a larger number of experimental animals. The findings of this study will contribute to more accurate experimental design of toxicity tests when using biomarkers. This should encourage other researchers to increase their effort to know the physiological state of their test organisms., (Copyright © 2011 Elsevier Inc. All rights reserved.)

A number of reports on potential toxicity of nanoparticles are available, but there is still a lack of knowledge concerning bioaccumulation. The aim of this work was to investigate how different sources of zinc, such as uncoated and unmodified ZnO nanoparticles, ZnCl(2) in solution, and macropowder ZnO influence the bioaccumulation of this metal in the terrestrial isopod Porcellio scaber. After exposure to different sources of Zn in the diet, the amount of assimilated Zn in whole body, the efficiency of zinc assimilation, and bioaccumulation factors (BAFs) were assessed. The bioaccumulation potential of Zn was found to be the same regardless of Zn source. The amount of assimilated Zn and BAF were dose-dependent, and Zn assimilation efficiency was independent of exposure concentrations. The Zn assimilation capacity was found to be up to 16% of ingested Zn. It is known that as much as approximately 20% of Zn can be accreted from ZnO particles by dissolution. We conclude that bioaccumulation of Zn in isopods exposed to particulate ZnO depends most probably on Zn dissolution from ZnO particles and not on bioaccumulation of particulate ZnO.

New products of nanotechnologies, including nanoparticles, need to be assessed according to their biological reactivity and toxic potential. Given the large number of diverse nanomaterials, a tiered approach is favoured. The aim of our work presented here is to elaborate an in vivo assay with terrestrial invertebrates (Porcellio scaber), which could serve as a first step of hazard identification of nanoparticles. We adapted the widely used acridine orange/ethidium bromide (AO/EB) assay to be applicable for cell membrane stability assessment of entire organ where the animal was exposed in vivo. The digestive glands (hepatopancreas) of terrestrial isopods were taken as a model test system. The assay was validated with Cu(NO(3))(2) and surfactants. The results showed that all tested nanoparticles, i.e. nanosized TiO(2), nanosized ZnO and fullerenes (C(60)) have cell membrane destabilization potential. As expected, C(60) is the most biologically potent. The AO/EB in vivo assay proved to be fast because response is recorded after 30 min of exposure, relatively simple because digestive glands are inspected immediately after isolation from exposed animals and promising approach because different types of nanoparticles could be tested for their biological potential. This assay provides data for the identification of hazardous potential of nanoparticles before subsequent steps in a tiered approach are decided.

The aim of this paper is to provide an insight into the past, current and future state of education in toxicology. Curricula in toxicology and environmental sciences at some European Universities are presented and discussed in the light of the 1999 Bologna reform. One of the key points in the Bologna reform is establishing a European Credit Transfer System (ECTS). An ECTS credit should reflect the expected learning outcomes of a student and should also be prospective of the value of lifelong learning. The Bologna reform encourages cooperation among institutions of higher education, recognition of variety of types of formal and informal education, implementation of lifelong education, joint programs and mobility. The other important point of Bologna reform is introduction of a new Master's level curricula for professional specialization which should be adapted to the specific skills demanded by the society. Such Master's level with competence based curriculum is a turning point in organizing toxicological education, which is necessary for the development of the profession. In addition, promotion of European cooperation and quality assurance within higher education could significantly benefit to toxicological education. The new Bologna education scheme appears to be suitable to direct toxicology toward a modern interdisciplinary profession capable of dealing with present and future challenges.

We report the use of a focused ion beam/scanning electron microscope (FIB/SEM) for simultaneous investigation of digestive gland epithelium gross morphology and ultrastructure of multilamellar intracellular structures. Digestive glands of a terrestrial isopod (Porcellio scaber, Isopoda, Crustacea) were examined by FIB/SEM and by transmission electron microscopy (TEM). The results obtained by FIB/SEM and by TEM are comparable and complementary. The FIB/SEM shows the same ultrastructural complexity of multilamellar intracellular structures as indicated by TEM. The term lamellar bodies was used for the multillamellar structures in the digestive glands of P. scaber due to their structural similarity to the lamellar bodies found in vertebrate lungs. Lamellar bodies in digestive glands of different animals vary in their abundance, and number as well as the thickness of concentric lamellae per lamellar body. FIB/SEM revealed a connection between digestive gland gross morphological features and the structure of lamellar bodies. Serial slicing and imaging of cells enables easy identification of the contact between a lamellar body and a lipid droplet. There are frequent reports of multilamellar intracellular structures in different vertebrate as well as invertebrate cells, but laminated cellular structures are still poorly known. The FIB/SEM can significantly contribute to the structural knowledge and is always recommended when a link between gross morphology and ultrastructure is investigated, especially when cells or cellular inclusions have a dynamic nature due to normal, stressed or pathological conditions.

The province of Guizhou in Southwestern China is currently one of the world's most important mercury production areas. Emissions of mercury from the province to the global atmosphere have been estimated to be approximately 12% of the world total anthropogenic emissions. The main objective of this study was to assess the level of contamination with Hg in two geographical areas of Guizhou province. Mercury pollution in the areas concerned originates from mercury mining and ore processing in the area of Wanshan, while in the area of Quingzhen mercury pollution originates from the chemical industry discharging Hg through wastewaters and emissions to the atmosphere due to coal burning for electricity production. The results of this study confirmed high contamination with Hg in soil, sediments and rice in the Hg mining area in Wanshan. High levels of Hg in soil and rice were also found in the vicinity of the chemical plant in Quingzhen. The concentrations of Hg decreased with distance from the main sources of pollution considerably. The general conclusion is that Hg contamination in Wanshan is geographically more widespread, due to deposition and scavenging of Hg from contaminated air and deposition on land. In Quingzhen Hg contamination of soil is very high close to the chemical plant but the levels reach background concentrations at a distance of several km. Even though the major source of Hg in both areas is inorganic Hg, it was observed that active transformation of inorganic Hg to organic Hg species (MeHg) takes place in water, sediments and soils. The concentration of Hg in rice grains can reach up to 569 microg/kg of total Hg of which 145 microg/kg was in MeHg form. The percentage of Hg as MeHg varied from 5 to 83%. The concentrations of selenium can reach up to 16 mg/kg in soil and up to 1 mg/g in rice. A correlation exists between the concentration of Se in soil and rice, indicating that a portion of Se is bioavailable to plants. No correlation between Hg and Se in rice was found. Exposure of the local population to Hg may occur due to inhalation of Hg present in air (in particular in Hg mining area) and consumption of Hg contaminated food (in particular rice and fish) and water. Comparison of intake through these different routes showed that the values of Hg considerably exceed the USA EPA Reference Concentration (RfC) for chronic Hg exposure (RfC is 0.0004 mg/m(3)) close to the emission sources. Intake of Hg through food consumption, particularly rice and fish, is also an important route of Hg exposure in study area. In general, it can be concluded that the population mostly at risk is located in the vicinity of smelting facilities, mining activities and close to the waste disposal sites in the wider area of Wanshan. In order to assess the real level of contamination in the local population, it is recommended that biomonitoring should be performed, including Hg and MeHg measurements in hair, blood and urine samples.

The biological cycle of mercury in the terrestrial isopod Porcellio scaber was investigated. Testing the possibility of in vivo Hg(2+) methylation was divided into two methodologically different parts. Firstly, concentrations of total mercury and MeHg in isopods P. scaber and their environment from a Hg-unpolluted area were measured by the use of validated methods (CV AAS, CV AFS). The data obtained show that the percentage of MeHg in leaves, soil and faeces was less than 1%. In contrast, the percentage of MeHg in gut and hepatopancreas was increased to 14 and 77%, respectively, indicating methylation of Hg(2+) in the gut and its further accumulation in glands. To confirm this assumption, the second methodology was applied-a radiotracer technique with 203Hg(2+) of high specific activity. There are few radiotracer techniques for Hg-methylation assays; for our work we chose the method of Czuba et al. which includes alkaline leaching of Hg species, their extraction into dithizone-toluene, followed by specific separation of Hg dithizonates by thin-layer chromatography and gamma counting. All steps of the analytical protocol were checked and optimised by the use of aqueous solutions of 203Hg(2+) and Me(203)Hg(+). The most important finding was that cleaning-up the extract through a florisil column is not appropriate, because the column retains different percentages of Hg(2+) and MeHg(+) and consequently affects the accuracy of the final result. This optimised protocol was then applied to Hg transformation studies in the terrestrial isopod P. scaber. Leaching Hg species from P. scaber fed with 203Hg(2+) or Me(203)Hg(+) dosed food was completely efficient only at elevated temperatures. Preliminary results of methylation/demethlytion studies are rather variable but they show that both processes (Hg(2+)<-->MeHg(+)) take place in the isopod P. scaber. Additionally, an assessment of the mass balance of Hg in isopods P. scaber exposed to 203Hg(2+) indicates that volatile Hg species are also formed.