Soil Invertebrate Enchytraeus Crypticus Research Articles

Environmental hazards of nanopesticides to non-target soil species - commercial nanoformulation versus its active substance (Karate Zeon® and lambda-cyhalothrin)

Nanopesticides (Npes) carry the potential of increased efficacy while reducing application rates, hence increasing agricultural productivity in a more sustainable way. However, given its novelty, the environmental risk assessment of these advanced materials is mostly absent. In the present study we investigated the ecotoxicity of a commercial insecticide, with reported nanofeatures, Karate Zeon®, and compared it to its active substance lambda-cyhalothrin. It is hypothesised that the use of the nanopesticide Karate Zeon® poses lower risk to enchytraeids than its active substance. The standard non-target soil invertebrate Enchytraeus crypticus was used, and exposure was done in LUFA 2.2 soil in 4 tests (endpoints: days): avoidance test [avoidance behaviour: 2 days], OECD standard reproduction test [survival, reproduction plus adults' size: 28 days] and its extension [total number organisms: 56 days], and Full Life Cycle (FLC) test [hatching and juveniles' size: 13 days; survival, reproduction and adults' size: 46 days]. Results showed that enchytraeids did not avoid Karate Zeon® nor its active substance lambda-cyhalothrin, which could be due to neurotoxicity. There was no indication of increased toxicity with prolonged exposure (46, 56d) compared to the standard (28d) for neither of the materials, being overall equally toxic in terms of hatching, survival, and reproduction. The FLCt results indicated that the juvenile stage was the most sensitive, resulting in higher toxicity for the adult animals when exposed from the cocoon stage. Although toxicity was similar between Karate Zeon and lambda-cyhalothrin, different patterns of uptake and elimination cannot be excluded. The benefits of using Karate Zeon will rely on reduced application rates.

Toxicokinetics and toxicodynamics of chromium in the soil invertebrate Enchytraeus crypticus (Oligochaeta)

Chromium emissions led to increased concentrations in soil, where it can affect soil organisms to relevant levels. With the aim of better understanding the effects of Cr throughout time, its toxicokinetics-toxicodynamics (TKTD) were evaluated in the soil model organism Enchytraeus crypticus to assess the development of internal concentrations and consequent toxic effects. To achieve this goal, organisms were exposed in LUFA 2.2 soil spiked with increasing CrCl3 concentrations. During the 21-day exposure period, survival, internal concentrations, and reproduction were evaluated at several time points up to 21 days. Uptake and elimination rate constants were 0.0044 kg soil/kg organism/day and 0.023 per day, respectively. Internal Cr concentrations increased with time, generally reaching equilibrium within 14 days with an estimated LC50inter (based on internal metal concentrations) of 57.7 mg Cr/kg body DW. Internal Cr concentrations were regulated by the organisms up to exposure to 360 mg Cr/kg soil DW, where the elimination rate was highest, but at 546 mg Cr/kg soil DW the animals were no longer able to eliminate Cr, and the internal concentrations were well above the estimated LC50inter. At day 21, exposure to 546 mg Cr/kg soil DW significantly reduced survival by 23 %, while reproduction EC50 was 344 mg Cr/kg soil DW. This study highlights the advantages of using a TKTD approach to understand the development of internal metal concentrations in time and relate it to the phenotypical effects observed. Toxicity is better understood when also taking into account time and not just exposure concentration alone.

Toxicokinetics and toxicodynamics of copper and cadmium in the soil invertebrate Enchytraeus crypticus (Oligochaeta)

The aim of this study was to evaluate the toxicokinetics-toxicodynamics (TKTD) of Cu and Cd in the soil model organism Enchytraeus crypticus, and assess the development of internal effect concentrations over time. Animals were exposed in LUFA 2.2 soil spiked with increasing concentrations of Cu and Cd. Survival, reproduction and internal metal concentrations in the animals were evaluated at different points in time over a period of 21 days. Internal concentrations increased with time, for Cu reaching a steady state after c. 10 days, except for the highest test concentration, and for Cd continuing to increase after 21 days. Applying a one-compartment model to all data together, estimated uptake and elimination rate constants for Cu and Cd were 0.08 and 0.45 kg soil/kg organism/day and 0.4 and 0.04 per day, respectively. Median lethal concentrations, based on total soil concentrations, decreased with time for Cu and did not reach a steady state level, but they did not change with time for Cd. The LC50inter (based on internal concentrations) was 75 mg Cu/kg body DW and > 800 mg Cd/kg body weight. Animals were able to regulate Cu internal concentrations, keeping them low, while for Cd internal concentrations continued to increase showing lack of regulation and also the importance of exposure time. This study highlights the advantages of using a TKTD approach to understand the relation between organism survival and internal Cu or Cd concentrations over time.

Toxicokinetics of metals in the soil invertebrate Enchytraeus crypticus exposed to field-contaminated soils from a mining area

Toxicokinetics may help assessing the risk of metal-contaminated soils by quantifying the development of internal metal concentrations in organisms over time. This study assessed the toxicokinetics in Enchytraeus crypticus of non-essential (Pb and Cd) and essential elements (Zn and Cu) in metal-contaminated field soils from a mining area, containing 3.49–24.3 mg Cd/kg dry soil, 433–1416 mg Pb/kg dry soil, 15.7–44.9 mg Cu/kg dry soil and 1718–6050 mg Zn/kg dry soil. Three different uptake-elimination patterns in E. crypticus were found. Both essential elements (Zn and Cu) showed fast increasing internal concentrations reaching equilibrium within 2 d in the uptake phase, without hardly any elimination after transfer to clean soil. The non-essential Cd showed a slow linear accumulation and excretion with body concentrations not reaching steady state within 21 d. Internal Pb concentrations, however, reached equilibrium within 7 d in the uptake phase. Longer exposure times in ecotoxicological tests, therefore, are required for elements like Cd. Porewater pH and dissolved organic carbon (DOC) levels were the dominant factors controlling Cd uptake from the test soils. The 21-d body Cd and Pb concentrations were best explained from 0.01 M CaCl2-extractable soil concentrations. Steady-state Cu and Zn body concentrations were independent of soil exposure concentrations. Bioaccumulation factors (BAF) were low for Pb (<0.1 kgsoil/kgworm), but high for Cd at 1.78–24.3 kgsoil/kgworm, suggesting a potential risk of Cd biomagnification in the terrestrial food chain of the mining area ecosystem.

Biochar and urban solid refuse ameliorate the inhospitality of acidic mine tailings and foster effective spontaneous plant colonization under semiarid climate

Phytomanagement is considered a suitable option in line with nature-based solutions to reduce environmental risks associated to metal(loid) mine tailings. We aimed at assessing the effectiveness of biochar from pruning trees combined with compost from urban solid refuse (USR) to ameliorate the conditions of barren acidic (pH ~5.5) metal(loid) mine tailing soils (total concentrations in mg kg−1: As ~220, Cd ~40, Mn ~1800, Pb ~5300 and Zn ~8600) from Mediterranean semiarid areas and promote spontaneous plant colonization. Two months after amendment addition were enough to observe improvements in chemical and physico-chemical tailing soil properties (reduced acidity, salinity and water-soluble metals and increased organic carbon and nutrients content), which resulted in lowered ecotoxicity for the soil invertebrate Enchytraeus crypticus. Recalcitrant organic carbon provided by biochar remained in soil whereas labile organic compounds provided by USR were consumed over time. These improvements were consistent for at least one year and led to lower bulk density, higher water retention capacity and higher scores for microbial/functional-related parameters in the amended tailing soil. Spontaneous growth of native vegetation was favored with amendment addition, but adult plants of remarkable size were only found after three years. This highlights the existence of a time-lag between the positive effects of the amendment on tailing soil properties being observed and these improvements being translated into effective spontaneous plant colonization.

Embryotoxicity of silver nanomaterials (Ag NM300k) in the soil invertebrate Enchytraeus crypticus – Functional assay detects Ca channels shutdown

Despite that silver (Ag) is among the most studied nanomaterials (NM) in environmental species and Ag’s embryotoxicity is well known, there are no studies on Ag NMs embryotoxicity in soil invertebrates. Previous Full Life Cycle (FLC) studies in Enchytraeus crypticus, a standard soil invertebrate, showed that Ag materials decreased hatching success, which was confirmed to be a hatching delay effect for silver nitrate (AgNO3) and mortality for Ag NM300K. In the present study, we aimed to investigate if the impact of Ag takes place during the embryonic development, using histology and immunohistochemistry. E. crypticus cocoons were exposed to a range of concentrations of Ag NM300K (0–10–20–60–115 mg Ag/kg) and AgNO3 (0–20–45–60–96 mg Ag/kg) in LUFA 2.2 soil, in an embryotoxicity test, being sampled at days 1, 2, 3 and 6 (3, 4, 5 and 7 days after cocoon laying). Measured endpoints included the number of embryonic structures, expression of transferrin receptor (TfR) and L type calcium channels (LTCC) through histological and immunohistochemistry analysis, respectively. Results confirmed that Ag materials affected the embryonic development, specifically at the blastula stage (day 3). The expression and localization of TfR in E. crypticus was shown in the teloblasts cells, although this transcytosis mechanism was not activated. Ag affected calcium (Ca) metabolism during embryonic development: for AgNO3, LTCC was initially activated, compensating the impact, for Ag NM300K, LTCC was not activated, hence no Ca balance, with irreversible consequences, i.e. terminated embryonic development. An Adverse Outcome Pathway was drafted, integrating the mechanisms here discovered with previous knowledge.

Toxicokinetics of copper and cadmium in the soil model Enchytraeus crypticus (Oligochaeta)

Toxicokinetics information is key to understanding the underlying intoxication processes, although this is often lacking. Hence, in the present study the toxicokinetics of copper (Cu) and cadmium (Cd) was assessed in the soil invertebrate Enchytraeus crypticus. The animals were exposed in LUFA 2.2 natural soil spiked to the estimated EC20 for reproduction effects in the Enchytraeid Reproduction Test (ERT), i.e. 80 mg Cu/kg soil Dry Weight (DW) and 20 mg Cd/kg soil DW. Tests followed the OECD guideline 317, including a 14-day uptake phase in spiked soil followed by 14 days elimination in clean soil, with samplings at days 0, 1, 2, 4, 7, 10, and 14. Exposure to Cu showed fast uptake, reaching a steady state after approx. 7 days, whereas for Cd, internal concentration increased and did not reach a clear steady state even after 14 days. When transferred to clean soil, Cu was rapidly eliminated returning to initial levels, while Cd-exposed animals still contained increased residue levels after 14 days. These differences in toxicokinetics have consequences for the toxicity and toxicodynamics and are indicative of the way essential and non-essential elements are handled by enchytraeids, likely also other soil invertebrates. This argues for the relevancy of longer exposure testing for elements like Cd compared to Cu, where phenotypical effects can well occur later at non-tested periods, e.g. after the 21 days’ duration of the standard ERT using E. crypticus.

Multigenerational Exposure to WCCo Nanomaterials-Epigenetics in the Soil Invertebrate Enchytraeus crypticus.

It has become clear how important it is to assess longer term effects of (nano) materials in the environment given the current evidence showing how epigenetics drives response mechanisms. Here we studied global DNA methylation in standard soil invertebrate Enchytraeus crypticus over 224 days when exposed to nanostructured tungsten carbide cobalt (WCCo nanomaterials (NMs)) and to cobalt (CoCl2) in a multigenerational experiment. In order to assess the transgenerational effect, we used a multigenerational (MG) test design consisting of four generations in spiked soil followed by two generations in clean soil. Results showed that MG exposure to WCCo NMs caused global DNA methylation to increase, which continued in unexposed generations and was associated with an increase in reproduction (phenotypic effect). In general, WCCo NMs caused more (and more consistent) methylation than CoCl2.

Protective effect of N-acetylcysteine on the toxicity of silver nanoparticles: Bioavailability and toxicokinetics in Enchytraeus crypticus.

We previously demonstrated that N-acetylcysteine (NAC) could reduce the toxicity of silver (Ag) materials (nanoparticles (NPs) and Ag nitrate) to the soil invertebrate Enchytraeus crypticus (Oligochaeta). It remains however, unclear whether the antitoxic mechanism of NAC was caused by NAC-Ag binding in the soil or inside the organisms. This study aimed at determining the bioavailability of Ag in the soil in a 21-day toxicity test as well as the Ag uptake and elimination kinetics in E. crypticus exposed to AgNPs in LUFA 2.2 standard soil amended with low (100 mg/kg dry soil) and high (600 mg/kg dry soil) NAC concentrations. The addition of NAC to the soil alleviated the toxicity of AgNPs by decreasing the internal Ag concentration of E. crypticus in a dose-dependent manner. Indeed, NAC reduced the binding of Ag to the soil, which probably was due to the formation of soluble but biologically unavailable Ag-cysteine complexes. The reduced Ag uptake in the enchytraeids was explained from an increased elimination at high NAC levels. These findings reinforce the view that metal complexing-compounds like NAC play a key role in the modulation of AgNP toxicity and bioavailability in terrestrial environments. Further, it may inform on the potential of NAC as a remediation solution for Ag or other metal-contaminated soils.

Do combined nanoscale polystyrene and tetracycline impact on the incidence of resistance genes and microbial community disturbance in Enchytraeus crypticus?

It has been proved that nanoplastics can effectively adsorb pollutants and thus influence their behavior and availability. The combined toxic effects of nanoplastic and its adsorbed pollutant on the soil fauna are still not well known. We used high-throughput quantitative PCR to explore the effects of oral nanoscale polystyrene and tetracycline exposure on antibiotic resistance genes in the soil invertebrate Enchytraeus crypticus, and used bacterial 16S rRNA gene amplification sequencing to examine the response of the microbiome of E. crypticus. After 14 days of tetracycline and nanoscale polystyrene exposure, we terminated exposure and monitored the restoration of ARGs and microbiome in the E. crypticus. Results showed that the number of ARGs, especially macrolide-lincosamide-streptogramin B (MLSB), tetracycline ARGs, as well as multidrug ARGs, increased with exposure to nanoscale polystyrene and tetracycline. The abundance of Aminoglycoside and Beta_Lactamase ARGs in E. crypticus also significantly increased. The exposure significantly perturbed the abundance of families Microbacteriaceae, Streptococcaceae, Enterobacteriaceae, Rhodocyclaceae and Sphinomonadaceae. After terminating exposure for 14 days, the diversity and abundance of ARGs were not completely restored, while the microbiome was not permanently changed but reversibly impacted.

The toxicity of silver nanomaterials (NM 300K) is reduced when combined with N-Acetylcysteine: Hazard assessment on Enchytraeus crypticus

The widespread production and use of silver nanomaterials (AgNMs) in consumer and medical products have been raising environmental concerns. Once in the environment, the soil is one of the major sinks of AgNMs due to e.g. sewage sludge applications, and invertebrates are directly exposed. In this study, we investigate the potential of N-acetylcysteine (NAC) to reduce the toxic effects of Ag NM300 K (and AgNO3) on the soil invertebrate Enchytraeus crypticus. Ag NM300 K induces mortality, reproduction impairment, and avoidance. The addition of NAC to the soil showed a remarkable reduction in the toxicity of Ag, indicating that NAC can act as a detoxifying agent for terrestrial organisms exposed to Ag materials. That the reduction in toxicity likely is caused by thiol groups, was confirmed by GSH and GSSH studies. Identifying the mechanisms and hence alternatives that allow the recovery of contaminated soils is an important mitigation measure to promote environmental safety and reduce the associated risks to human health. Further, it may inform on strategies to implement in safe-by-design industry development.

Does soil CuO nanoparticles pollution alter the gut microbiota and resistome of Enchytraeus crypticus?

Growing evidence suggests that metallic oxide nanoparticles can pose a severe risk to the health of invertebrates. Previous attention has been mostly paid to the effects of metallic oxide nanoparticles on the survival, growth and physiology of animals. In comparison, the effects on gut microbiota and incidence of antibiotic resistance genes (ARGs) in soil fauna remain poorly understood. We conducted a microcosm study to explore the responses of the non-target soil invertebrate Enchytraeus crypticus gut microbiota and resistomes to copper oxide nanoparticles (CuO NPs) and copper nitrate by using bacterial 16S rRNA gene amplicons sequencing and high throughput quantitative PCR. The results showed that exposure to Cu2+ resulted in higher bioaccumulation (P < 0.05) and lower body weight and reproduction (P < 0.05) of Enchytraeus crypticus than exposure to CuO NPs. Nevertheless, exposure to CuO NPs for 21 days markedly increased the alpha-diversity of the gut microbiota of Enchytraeus crypticus (P < 0.05) and shifted the gut microbial communities, with a significant decline in the relative abundance of the phylum Planctomycetes (from 37.26% to 19.80%, P < 0.05) and a significant elevation in the relative abundance of the phyla Bacteroidetes, Firmicutes and Acidobacteria (P < 0.05). The number of detected ARGs in the Enchytraeus crypticus gut significantly decreased from 45 in the Control treatment to 16 in the Cu(NO3)2 treatment and 20 in the CuO NPs treatment. The abundance of ARGs in the Enchytraeus crypticus gut were also significantly decreased to 38.48% when exposure to Cu(NO3)2 and 44.90% when exposure to CuO NPs (P < 0.05) compared with the controls. These results extend our understanding of the effects of metallic oxide nanoparticles on the gut microbiota and resistome of soil invertebrates.

Graphene-Based Nanomaterials in Soil: Ecotoxicity Assessment Using Enchytraeus crypticus Reduced Full Life Cycle.

Graphene-based nanomaterials (GBNs) possess unique physicochemical properties, allowing a wide range of applications in physical, chemical, and biomedical fields. Although GBNs are broadly used, information about their adverse effects on ecosystem health, especially in the terrestrial environment, is limited. Therefore, this study aims to assess the toxicity of two commonly used derivatives of GBNs, graphene oxide (GO) and reduced graphene oxide (rGO), in the soil invertebrate Enchytraeus crypticus using a reduced full life cycle test. At higher exposure concentrations, GO induced high mortality and severe impairment in the reproduction rate, while rGO showed little adverse effect up to 1000 mg/kg. Collectively, our body of results suggests that the degree of oxidation of GO correlates with their toxic effects on E. crypticus, which argues against generalization on GBNs ecotoxicity. Identifying the key factors affecting the toxicity of GBNs, including ecotoxicity, is urgent for the design of safe GBNs for commercial purposes.

Effect of ageing and chemical form on the bioavailability and toxicity of Pb to the survival and reproduction of the soil invertebrate Enchytraeus crypticus

This study investigated the effect of ageing on the bioavailability and toxicity of lead nitrate (Pb(NO3)2) and lead oxide (PbO) to Enchytraeus crypticus in LUFA 2.2 natural soil. The potworms were exposed after 2 weeks pre-incubation and after ageing the spiked soils for 3, 6, 12 and 18 months. Survival and reproduction after 21 d exposure were related to total, 0.01 M CaCl2-extractable and porewater Pb concentrations in the soil and internal Pb concentrations in the surviving animals. Pb concentration in pore water showed little change during ageing for Pb(NO3)2 but increased strongly for PbO-spiked soils. During ageing, toxicity of Pb(NO3)2 did not change with LC50s and EC50s for the effect on enchytraeid survival and reproduction based on total soil Pb concentrations being constant at 523–619 and 89.8–99.4 mg Pb/kg dry soil, respectively. Toxicity of PbO, however, increased with LC50s and EC50s decreasing from 4830 to 1889 mg Pb/kg dry soil and from 151 to 97.5 mg Pb/kg dry soil, respectively. When related to internal Pb concentrations LC50s did not differ for both Pb forms at different ageing periods and were 73.4–78.7 mg Pb/kg dry body wt. Survival was better explained from internal Pb concentrations in the worms than from total or available Pb concentrations in the soil. Reproduction toxicity (EC50s) and Pb uptake in the worms however, were better explained from 0.01 M CaCl2-extractable Pb concentrations in the soil. The latter finding could provide a scientific basis for the ecological risk assessment of contaminated soils and the derivation of soil quality standards based on extractable concentrations.

Effect of percolation and chemical form on Pb bioavailability and toxicity to the soil invertebrate Enchytraeus crypticus in freshly spiked and aged soils

In standard terrestrial ecotoxicological tests, soils usually are spiked with highly soluble metal salts leading to overestimation of bioavailability and introducing counterions that may contribute to toxicity. Leaching is suggested as an efficient method to avoid the effects of the associated counterions. The present study aimed at investigating the effects of leaching on the bioavailability and toxicity of Pb(NO3)2 and PbO to the potworm Enchytraeus crypticus in LUFA 2.2 soil freshly spiked or after 18 months ageing. Percolation decreased porewater Pb concentrations as well as the toxicity of both Pb forms. The influence of percolation differed between the two Pb forms and between freshly spiked and aged soils. Percolation slightly increased LC50s based on total soil Pb concentrations for Pb(NO3)2, but not for PbO, and only affected Pb toxicity to enchytraeid reproduction in freshly spiked soils. The differences in Pb uptake in E. crypticus and toxicity between the two Pb forms as well as between different treatments could be minimized by relating them to 0.01 M CaCl2-extractable concentrations. In addition, body Pb concentrations could well explain enchytraeid survival across all soils and treatments, indicating its suitability as a good proxy for Pb toxicity in soil.

Exposure to tetracycline perturbs the microbiome of soil oligochaete Enchytraeus crypticus

Microbial symbiosis is essential for the normal development and growth of hosts. Past attention has mostly been paid to its effects on plants and vertebrates. The effects of environmental pressures such as antibiotics on the microbiome of soil fauna remain largely elusive. We used bacterial 16S rRNA gene high-throughput sequencing to examine the response of microbiome of soil invertebrate Enchytraeus crypticus to oral tetracycline exposure. After two-week exposure, tetracycline-free oat was used as food to monitor the restoration of E. crypticus microbiome. The results showed that Proteobacteria, Actinobacteria and Planctomycetes were the three dominant phyla in all samples, Rhizobiaceae and Kaistia were the most abundant family and genus in all samples, respectively. After 14 days tetracycline exposure, Planctomycetes declined dramatically from 33.05% to 3.28% (P = 0.016), but Actinobacteria elevated substantially from 2.47% to 23.65% (P = 0.004). The alpha-diversity of microbial community increased significantly after tetracycline exposure compared to the control (P = 0.014). Terminating tetracycline exposure led to the recovery of E. crypticus microbiome back to the background level within 14 days. Our results suggest that while tetracycline can disturb the microbiome in E. crypticus significantly, the effects of the antibiotic on E. crypticus microbiome may not be permanent but reversibly diminish after stopping exposure for a period of time. The results may contribute to extending our understanding of the effect of antibiotics on microbiome of soil invertebrates. CapsuleThe microbiome of E. crypticus exposed to tetracycline is perturbed and reversibly restored after terminating the exposure.

A combined toxicokinetics and toxicodynamics approach to investigate delayed lead toxicity in the soil invertebrate Enchytraeus crypticus

In a previous study, Pb toxicity was found to be delayed compared to Pb bioaccumulation in Enchytraeus crypticus. This study aimed at further investigating the acute and delayed onset of Pb toxicity in E. crypticus by using a combination of toxicokinetics and toxicodynamics approaches. Enchytraeids were exposed to different Pb concentrations (uptake phase) in natural LUFA 2.2 soil for different short-term exposure periods, followed by a 7-d elimination phase in clean soils. Body Pb concentrations and enchytraeid mortality were determined at different time intervals during both the exposure and the elimination phase. Pb uptake kinetics in E. crypticus were well described by a three-stage first-order model with an initial overshoot in body Pb concentrations. At higher exposure concentrations, Pb caused delayed enchytraeid mortality even following short-term exposure. LC50 based on body Pb concentrations appeared no good descriptor of delayed Pb toxicity in E. crypticus. Exposure time had a major impact on Pb bioaccumulation, toxicity and its delayed effects, which argues against relying on ecotoxicity tests for metal toxicity using a fix exposure duration. The presence of delayed toxic effects also suggests that post-exposure observations are necessary to avoid underestimation of metal toxicity.

Effect of soil properties on Pb bioavailability and toxicity to the soil invertebrate Enchytraeus crypticus

The present study investigated the bioavailability and toxicity of lead to the potworm Enchytraeus crypticus in six soils with different properties. Pb partitioning between the soil solution and solid phase was affected by soil organic matter (OM) content, cation exchange capacity (CEC) and water holding capacity (WHC). After 21 d exposure, Pb bioaccumulation in the enchytraeids was positively correlated with total soil Pb concentration. Bioaccumulation was best predicted by Pb availability (CaCl2-extractable and porewater Pb concentrations), and by the Ca concentration in pore water and the CEC of the soils. Toxicity varied greatly among soils, with LC50s and EC50reproductions based on total Pb concentrations ranging from 246 to >3092 and from 81 to 1008 mg Pb/kg dry soil, respectively. The variation in LC50s among soils was explained by differences in CaCl2-extractable Pb concentrations in soil and internal Pb concentrations in the animals. The differences in EC50reproductions could be explained from the CaCl2-extractable Pb concentrations in the soils. Although it was also correlated with CEC and porewater Ca concentration, pHCaCl2 was the dominating factor for predicting Pb toxicity based on total soil concentrations. This study demonstrates that soil properties, such as pH, CEC and Ca concentration in pore water, significantly affected the bioavailability and toxicity of Pb and therefore should be taken into account when assessing the ecological risk of metals in contaminated soils.

Multigenerational effects of copper nanomaterials (CuONMs) are different of those of CuCl2: exposure in the soil invertebrate Enchytraeus crypticus

Nanomaterials (NMs) are recommended to be tested in longer term exposures. Multigenerational (MG) studies are scarce and particularly important because effects can be transferred to the next generation. The current risk assessment framework does not include MG effects and this is a caveat for persistent materials. Here, the effects of copper NMs (CuONMs) and copper salt (CuCl2) were assessed in a MG exposure (4 generations in spiked soil + 2 generations in clean soil, F1 to F7 generations in total), with the standard soil model Enchytraeus crypticus, using relevant reproduction test effect concentrations (EC10, EC50), monitoring survival and reproduction. This represented ca. 1 year continuous exposure tests. MG effects varied with effect concentration and test materials: CuONMs caused increased toxicity for EC10 exposed organisms (EC50 did not change), and transfer to clean media reset effects, whereas CuCl2 reduced toxicity for EC10 and EC50, but the transfer to clean media “revived” the initial effects, i.e. close to EC50 levels in F7. Clearly CuONMs and CuCl2 cause different mechanisms of toxicity or response in the long term, not predictable based on short term or one generation studies. The present contributes for the improvement of risk assessment, adding important information for the long term exposure and effects.

Time-dependent uptake and toxicity of nickel to Enchytraeus crypticus in the presence of humic acid and fulvic acid.

The present study aimed to investigate the influence of different fractions of dissolved organic carbon (DOC) on the uptake and toxicity of nickel (Ni) in the soil invertebrate Enchytraeus crypticus after different exposure times. The addition of DOC as humic acid or fulvic acid significantly reduced Ni uptake by E. crypticus in the soil-solution test system. Median lethal effect concentrations were calculated based on total dissolved Ni concentrations (LC50[Ni]), free Ni ion activity (LC50{Ni2+ }), and Ni body concentrations (LC50Body-Ni ). The LC50[Ni] values increased with increasing DOC levels and decreased with exposure time (4, 7, and 10 d). Humic acid exerted a greater protective effect on Ni toxicity than fulvic acid, but the protective effects decreased with prolonged exposure time. The LC50{Ni2+ } values also decreased with exposure time but were almost constant with variation in DOC levels, indicating that the protective effect of DOC is mainly through complexation with free Ni ions to reduce Ni bioavailability. The LC50Body-Ni value was independent of DOC concentration and exposure time, with an estimated overall value of 22.1 µg/g dry weight. The present study shows that body concentration could serve as an effective indicator for predicting Ni toxicity with variations in the exposure environment (e.g., DOC) and exposure time. Environ Toxicol Chem 2017;36:3019-3027. © 2017 SETAC.