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Polycyclic aromatic hydrocarbons (PAHs) continue to cause environmental challenges as a result of their release into the environment by a great variety of anthropogenic activities and their accumulation in soils. Studies were conducted on the toxicological effect of the model PAH phenanthrene using the soil invertebrate model Enchytraeus crypticus at the individual, tissue, and molecular levels. Animals were exposed for 2 d and 21 d to phenanthrene concentrations corresponding to the (previously estimated) 3-wk effective concentration, 10% (EC10) and EC50 for effects on reproduction. Gene expression profiling did not reveal a typical phenanthrene-induced biotransformation signature, as it usually does in arthropods and vertebrates. Instead, after 2 d of exposure, only general metabolic processes were affected, such as translation and adenosine triphosphate synthesis-coupled electron transport. Histological sections of tissues of 2-d exposed animals showed no deviations from control. In contrast, after prolonged exposure of up to 21 d, histopathological effects were found: chloragogenous cells were highly vacuolated and hypertrophic. This was corroborated by differential expression of genes related to immune response and oxidative stress at the transcriptomic level. The data exemplify the complexity and species-specific features of PAH toxicity among soil invertebrate communities, which restricts read-across and extrapolation in the context of soil ecological risk assessment. Environ Toxicol Chem 2016;35:2713-2720. © 2016 SETAC [ABSTRACT FROM AUTHOR]

Supralittoral ecosystems are among the most challenging environments for soil organisms, particularly when salinity fluctuations are involved, frequently combined with the presence of contaminants as a result of intense anthropogenic activities. Knowledge of how salinity influences the effect of contaminants in supralittoral species is crucial for determining the safety factors required when extrapolating results from optimal laboratory conditions to these natural ecosystems. The present study therefore evaluated the effects of 2 metals (copper and cadmium) and 2 organic compounds (carbendazim and 4-nonylphenol) in the absence or presence of 15‰ NaCl in the potworm Enchytraeus albidus, a model organism for ecotoxicology studies commonly found in supralittoral ecosystems, The potworms had a higher reproduction in saline soil than in control soil. Moreover, the effects of copper and carbendazim on reproduction were smaller than when they were tested in nonsaline soil. Potworms exposed to nonsaline soils also had significantly higher tissue concentrations of metals, which partly explains the effects on reproduction. The influence of salinity on effects of 4-nonylphenol was, however, less clear; effects on survival decreased in saline soil, but effects on reproduction were highest in saline soil. The latter slightly correlated with tissue concentrations of the chemical. The present study provides the first evidence that soil salinity has a significant influence on the impact of contaminants evaluated with the enchytraeid reproduction test. Environ Toxicol Chem 2015;34:2159-2166. © 2015 SETAC [ABSTRACT FROM AUTHOR]

Abstract: Triclosan (TCS) is an antimicrobial agent which is used as a broad-spectrum bacteriostatic and found in personal care products, and due to this it is spread in the environment. Aquatic toxicity studies have been reported, but little data is available for its toxicity to terrestrial organisms. The main purpose of this study was to assess the toxicity of TCS in the terrestrial environment, using a battery of soil species, belonging to different taxonomic levels. The species included the invertebrates Eisenia andrei, Enchytraeus albidus and Folsomia candida and the plants Triticum aestivum (monocotyledonous) and Brassica rapa (dicotyledonous). For the invertebrate species the reproduction EC10s were 0.6 (E. andrei), 2 (E. albidus) and 7(F. candida) mg TCS/kg soil dry weight (DW). For plants'' emergence EC10s are 0.1 (B. rapa) and 142 (T. aestivum) TCS/kg soil DW. The calculated PNEC (Predicted No Effect Concentration) ranged between 0.0008 and 0.004mg TCS/kg (95% certainty) and between 0.04 and 0.2mg TCS/kg soil DW (50% certainty) when using the SSD (Species Sensitivity Distribution) approach; applying a safety factor to the lowest EC10 resulted in a PNEC of 0.06mg TCS/kg soil DW. [Copyright &y& Elsevier]

Abstract: Contamination problems are often characterized by complex mixtures of chemicals. There are two conceptual models usually used to evaluate patterns of mixture toxicity: Concentration Addition (CA) and Independent Action (IA). Deviations from these models as synergism, antagonism and dose dependency also occur. In the present study, single and mixture toxicity of atrazine, dimethoate, lindane, zinc and cadmium were tested in Porcellionides pruinosus and Enchytraeus albidus, using avoidance as test parameter. For both species patterns of antagonism were found when exposed to dimethoate and atrazine, synergism for lindane and dimethoate exposures (with the exception of lower doses in the isopod case study) and concentration addition for cadmium and zinc occurred, while the exposure to cadmium and dimethoate showed dissimilar patterns. This study highlights the importance of dose dependencies when testing chemical mixtures and that avoidance tests can also be used to asses the effects of mixture toxicity. [Copyright &y& Elsevier]

Abstract: Enchytraeids are typical inhabitants of many soils, contributing to vital processes of this environmental compartment. Indirectly they are involved in regulating the degradation of organic matter, as well as improving the pore structure of the soil. Due to their behaviour, they are able to avoid unfavourable environmental conditions. Avoidance tests with enchytraeids, initially developed with earthworms by several authors, are quick and easy to perform. With these tests a first assessment of the toxicity of a (contaminated or spiked) soil is possible in just 48h by using the reaction of the enchytraeids as measurement endpoint. In this period of time the organisms can choose between the control soil and the other soil (a contaminated or spiked or another soil with different physico-chemical properties). In the tests reported here, the enchytraeids were exposed to control soils spiked with the fungicides Benomyl and Carbendazim and the herbicide Phenmedipham. Several chemical concentrations were tested in order to evaluate the avoidance behaviour to toxic substances. In fact, often these short-term screening tests gave results showing avoidance at concentrations in a range similar to the acute test results but, higher than in chronic tests. Further tests are needed to decide whether the results gained in this study can be extrapolated to other chemicals. It is proposed to standardize the Enchytraeid Avoidance Test as it is currently done for the Earthworm Avoidance Test by the International Standard Organization (ISO). [Copyright &y& Elsevier]

Abstract: Enchytraeids are important inhabitants of a wide variety of soil types and suitable test organisms for environmental risk assessment through the determination of effects on survival and reproduction. In this study, the effect of pesticides with different modes of action is assessed in the standard test species Enchytraeus albidus. Main goals were: (a) to test the toxicity of a range of pesticides to E. albidus in a natural soil (LUFA 2.2), (b) to assess whether avoidance gave a prediction of effects on the survival and reproduction, and (c) to investigate if the organisms’ response to different toxics can be grouped into the respective chemical classes. Pesticides selected were the: (1) herbicides: phenmedipham and atrazine; (2) fungicides: carbendazim and pentachlorophenol; and (3) insecticides: dimethoate and lindane. All tested pesticides caused effects in the survival and reproduction of E. albidus and the compounds that showed a higher toxicity were carbendazim, dimethoate and atrazine. The effect concentrations were not chemical class dependent. In general, survival and reproduction showed similar response patterns. Avoidance behaviour showed trends of response similar to these other measured endpoints, but was less sensitive and more variable (data from a previous study). It was not possible to establish a clear correlation between survival, reproduction and avoidance endpoints. From an ecological point of view, avoidance tests are relevant but due to lower sensitivity and higher variability it is recommended that the enchytraeid avoidance test should not be used for risk assessment purposes. [Copyright &y& Elsevier]

Abstract Gene expression can vary with the organisms' life stage. It is known that embryos can be more sensitive to toxicant exposure, as previously demonstrated for Enchytraeus crypticus (Oligochaeta) exposed to cadmium (Cd), known to cause embryotoxicity and hatching delay. It was shown that Ca enters embryos via the L-type Ca channels in the cocoon membrane, this being affected in Cd exposed embryos (Cd-Ca competition is well-known). In the present study, the embryotoxic mechanisms of Cd were studied via high-throughput gene expression for E. crypticus. Cocoons (1–2 days old), instead of the adult organism, were exposed in Cd spiked LUFA 2.2 soil during 1 day. Results showed that Cd affected Ca homeostasis which is implicated in several other molecular processes. Several of the major modulators of Cd toxicity (e.g., impaired gene expression, cell cycle arrest, DNA and mitochondrial damage) were identified in the embryos showing its relevancy as a model in ecotoxicogenomics. The draft Adverse Outcome Pathway was improved. Previously was hypothesized that gene regulation mechanisms were activated to synthesize more Ca channel proteins – this was confirmed here. Further, novel evidences were that, besides the extracellular competition, Cd competes intracellularly which causes a reduction in Ca efflux, and potentiates Cd embryotoxicity. Graphical abstract Image 1 Highlights • E. crypticus embryos are more sensitive to Cd than adults. • Embryos' mechanisms were studied using high-throughput gene expression. • Major modulators of Cd toxicity were identified in the embryos. • Cd competes with Ca inside the cells (novel), besides extracellular (known). • Adverse Outcome Pathway (AOP) was improved. [ABSTRACT FROM AUTHOR]

Information on effects of silver nanoparticles on soil invertebrates, especially using long-term exposures, is scarce. In this study we investigated the effects of the reference Ag (NM300K) (compared to AgNO 3 ) using the full life cycle test (FLCt) of the soil invertebrate Enchytraeus crypticus . Results showed that effects were higher compared to the standard reproduction test, which is shorter and does not cover the FLC. Both Ag forms caused a reduction on hatching success, juvenile and adult survival and reproduction with similar ECx. Differences between AgNO 3 and Ag NM300K could be discriminated using the FLCt: AgNO 3 decreased hatching success was shown to be a delay in the process, whereas Ag NM300K caused irreversible effects during the same time frame. These effects may have occurred during the embryo development, hatching (inhibition) or survival of hatched juveniles. Ag NM300K caused non-monotonic concentration-response effect as observed by the high effect of the lowest concentration (20 mg kg-1). It is known that dispersion is higher at lower concentrations – this could explain the increased effect at low concentration. Non monotonic responses are well described in the literature, where effects of high cannot predict for low concentrations, hence special attention should be given for NMs low concentration effects. [ABSTRACT FROM AUTHOR]

The nano-form of copper (Cu-NPs) is already extensively used. In this paper the toxic effect of Cu in the worm Enchytraeus crypticus (Oligochaeta: Enchytraeidae) was assessed following exposure to (1) Cu-salt: freshly spiked soil with copper-nitrate, (2) Cu-NPs: freshly spiked soil with Cu nanoparticles (80 nm), and (3) Cu-field: historically Cu contaminated soil (80 years ago). Our main aims were to compare the three different exposure regimes and respective toxicity, and to determine how the oxidation state of the Cu and dissolution state of the particles differed. Characterization of in situ -exposure included identification of oxidation states with synchrotron generated X-ray absorption near-edge spectroscopy (XANES) analysis, activity of free Cu 2+ in soil-solution (Ion Selective Electrode), and the relative distribution of the labile Cu-fractions (Sequential Extraction). Freshly spiked Cu-salt was the most toxic for reproductive output of the worms, followed by Cu-NPs and then Cu-field. XANES indicated only one oxidation state (II) in Cu-salt and Cu-field soil, whereas in Cu-NPs soil it was present in all oxidation states (0, I and II). The partial oxidation of the Cu-NPs (in soil) was evident and with limited dissolution. [ABSTRACT FROM AUTHOR]

The existing standard enchytraeid reproduction test (ERT) concerns the assessment of effects on survival and reproduction. In the present study we optimized and propose an embryotoxicity test using Enchytraeus crypticus. Cadmium (Cd) was used as a test substance. Endpoints evaluated were embryo development, number of embryonic structures, Calcium (Ca) channels quantification and hatching success with macroscopic monitoring, histological and immunohistochemistry analysis. Results showed that Cd is embryotoxic for this species, causing a decrease in the hatching success (EC 50 = 3.1 mg/kg), a delay or disruption in formation of embryonic structures depending on concentrations (<5 mg Cd/kg or ⩾16 mg Cd/kg). Results from immunohistochemistry suggest a competitive binding between Cd and Ca for Ca channels, resulting in changes in Ca homeostasis. The use of the E. crypticus embryotoxicity test with the combination of histological and immunohistological tools provided a good option towards mechanistic information enhancing the importance of these tests to evaluate the hazard of chemicals and possible use in risk assessment. [ABSTRACT FROM AUTHOR]