Microplastics originated from agricultural mulching films affect enchytraeid multigeneration reproduction and soil properties.

Microplastics (MPs) are increasingly entering agricultural soils, often from the breakdown of agricultural plastics (e.g., mulching films). This study investigates the effects of realistic MPs from different mulching films: two conventional polyethylene (PE-1 and PE-2) and two biodegradable (starch-blended polybutylene adipate co-terephthalate; PBAT-BD-1 and PBAT-BD-2). MPs were mixed into Lufa 2.2 soil at a concentration range from 0.005 % to 5 % (w/w dry soil), wide enough to reflect both realistic environmental levels and "worst-case scenarios". Effects on Enchytraeus crypticus reproduction over two generations and six important soil properties were studied. PBAT MPs notably reduced enchytraeid reproduction in the F0 generation, with a maximum decrease of 35.5 ± 9.6 % at 0.5 % concentration. F1 generation was unaffected by PBAT contamination. PE MPs had a more substantial reproductive impact, with up to a 55.3 ± 9.7 % decrease at 5 % PE-1 concentration compared to the control, showing a dose-related effect except for 1 %. Both MP types also significantly affected soil water holding capacity, pH, and total carbon. Other soil properties remained unaffected. Our results highlight the potential negative impacts of MPs originating from real agricultural plastics on soil health and raise concerns about the role of agricultural plastics in sustainable agriculture and food safety.

Foliar Exposure of Deuterium Stable Isotope-Labeled Nanoplastics to Lettuce: Quantitative Determination of Foliar Uptake, Transport, and Trophic Transfer in a Terrestrial Food Chain.

Nanoplastics (NPs) are widely detected in the atmosphere and are likely to be deposited on plant leaves. However, our understanding of their foliar uptake, translocation, and trophic transfer profiles is limited due to a lack of quantitative analytical tools to effectively probe mechanisms of action. Here, using synthesized deuterium (2H) stable isotope-labeled polystyrene nanoplastics (2H-PSNPs), the foliar accumulation and translocation of NPs in lettuce and the dynamics of NP transfer along a lettuce-snail terrestrial food chain were investigated. Raman imaging and scanning electron microscopy demonstrated that foliar-applied NPs aggregated on the leaf surface, entered the mesophyll tissue via the stomatal pathway, and eventually translocated to root tissues. Quantitative analysis showed that increasing levels of foliar exposure to 2H-PSNPs (0.1, 1, and 5 mg/L in spray solutions, equivalent to receiving 0.15, 1.5, and 7.5 µg/d of NPs per plant) enhanced NP accumulation in leaves, with concentrations ranging from 0.73 to 15.6 µg/g (dw), but only limited translocation (<5%) to roots. After feeding on 5 mg/L 2H-PSNP-contaminated lettuce leaves for 14 days, snails accumulated NPs at 0.33 to 10.7 µg/kg (dw), with an overall kinetic trophic transfer factor of 0.45, demonstrating trophic dilution in this food chain. The reduced ingestion rate of 3.18 mg/g/day in exposed snails compared to 6.43 mg/g/day can be attributed to the accumulation of 2H-PSNPs and elevated levels of chemical defense metabolites in the lettuce leaves, which decreased the palatability for snails and disrupted their digestive function. This study provides critical quantitative information on the characteristics of airborne NP bioaccumulation and the associated risks to terrestrial food chains.

Multigenerational toxicity of microplastics derived from two types of agricultural mulching films to Folsomia candida.

Degradation and fragmentation of mulching films represents an increasing source of microplastics (MPs, plastic particles 1 µm to 5 mm in size) to agricultural soils. MPs have been shown to affect many soil invertebrates, including springtails. However, these studies typically use test materials representing less environmentally relevant particle types, such as pristine uniform MPs, which do not represent the large range of particle sizes and morphologies found in the field. This study aimed at providing insight into the adverse effects of MPs originating from agricultural mulching films, by using artificially aged MPs derived from both biodegradable (starch-polybutadiene adipate terephthalate (PBAT)) blend, as well as conventional (linear low-density polyethylene (LLDPE)) plastic polymers. The soil dwelling springtail Folsomia candida was exposed to these MPs for five generations in order to elucidate population effects due to possible reproduction toxicity, endocrine disruption, mutagenesis or developmental toxicity. F. candida were exposed to 0, 0.0016, 0.008, 0.04, 0.2, 1, 2, 3, 4 and 5 % (w/w dry soil) MPs in Lufa 2.2 soil, which includes concentrations within the range of environmental relevance. Juveniles produced at each concentration were transferred to the next generation, with the parental, F2 and F4 generations being exposed for four weeks and F1 and F3 generations for five weeks. No concentration-dependent effects on F. candida survival or reproduction were observed in exposures to either of the MPs, in any of the generations. These results suggest that the particular MPs used in this study, derived from mulching films used on agricultural soils, may not be potent toxicants to F. candida, even after long-term exposure and at elevated concentrations.

Field-realistic doses of the neonicotinoid acetamiprid impact natural soil arthropod community diversity and structure.

The neonicotinoid acetamiprid is used as a foliar insecticide spray, which results in direct exposure of a wide variety of soil organisms. Laboratory testing indicated that acetamiprid is toxic to the Collembola (springtails) species Folsomia candida, while Acari (mites) seem relatively insensitive to neonicotinoids. Since such opposing effects on different soil arthropods might imbalance natural arthropod communities, this study determined: (i) if field-realistic doses of acetamiprid affect the abundance and diversity in soil arthropod communities, and (ii) whether these potential effects are short-term or persist after degradation of acetamiprid. Intact soil cores collected from an untreated grassland field were placed in the mesocosm set up 'CLIMECS', and the naturally sourced communities were exposed to a control and increasing field-realistic doses of acetamiprid (i.e. 0, 0.05, 0.2, 0.8 mg a.s./kg dry soil). Before and 7 and 54 days after spraying the insecticide, the abundance of mites and springtails and springtail diversity were assessed. Springtail and mite abundances were similar at the start of the experiment, but springtail abundance was significantly lowered while mite abundance increased shortly after exposure to increasing doses of acetamiprid. At the highest dose, springtail numbers decreased by 53% on average while the number of mites increased by 26%. This effect was no longer visible after 54 days, suggesting recovery of the community as a whole reflected by observed changes in community dissimilarity: shortly after application springtail communities clearly diverged from the control in terms of species composition, while communities converged again in the long-term. With our results, we are the first to show that field-realistic applications of N-nitroguanidine neonicotinoids can significantly impact natural soil fauna communities, which might have implications for soil ecosystem functioning.

Thermal adaptation affects the temperature-dependent toxicity of the insecticide imidacloprid to soil invertebrates.

Terrestrial ectotherms are vulnerable to climate change since their biological rates depend on the ambient temperature. As temperature may interact with toxicant exposure, climate change may cause unpredictable responses to toxic stress. A population's thermal adaptation will impact its response to temperature change, but also to interactive effects from temperature and toxicants, but these effects are still not fully understood. Here, we assessed the combined effects of exposure to the insecticide imidacloprid across the temperatures 10-25 °C of two populations of the Collembola Hypogastrura viatica (Tullberg, 1872), by determining their responses in multiple life history traits. The con-specific populations differ considerably in thermal adaptations; one (arctic) is a temperature generalist, while the other (temperate) is a warm-adapted specialist. For both populations, the sub-lethal concentrations of imidacloprid became lethal with increasing temperature. Although the thermal maximum is higher for the warm-adapted population, the reduction in survival was stronger. Growth was reduced by imidacloprid in a temperature-dependent manner, but only at the adult life stage. The decrease in adult body size combined with the absence of an effect on the age at first reproduction suggests a selection on the timing of maturation. Egg production was reduced by imidacloprid in both populations, but the negative effect was only dependent on temperature in the warm-adapted population, with no effect at 10 °C, and decreases of 41 % at 15 °C, and 74 % at 20 °C. For several key traits, the population best adapted to utilize high temperatures was also the most sensitive to toxic stress at higher temperatures. It could be that by allocating more energy to faster growth, development, and reproduction at higher temperatures, the population had less energy for maintenance, making it more sensitive to toxic stress. Our findings demonstrate the need to take into account a population's thermal adaptation when assessing the interactive effects between temperature and other stressors.

Pioneer plants enhance soil multifunctionality by reshaping underground multitrophic community during natural succession of an abandoned rare earth mine tailing.

Spontaneous natural succession in metal mine tailings is fundamental to the rehabilitation of bare tailing. Here, an abandoned rare earth element (REE) mine tailing with spontaneous colonisation by pioneer plants with different functional traits was selected. Soil nutrient cycling, fertility, organic matter decomposition as well as underground organismal communities and their multitrophic networks were investigated. Compared with the bare tailing, the colonisation with Lycopodium japonicum, Miscanthus sinensis, and Dicranopteris dichotoma increased soil multifunction by 222%, 293%, and 525%, respectively. This was accompanied by significant changes in soil bacterial and protistan community composition and increased soil multitrophic network complexity. Rhizospheres of different plant species showed distinct microbial community composition compared to that of bare tailing. Some WPS-2, Chloroflexi, and Chlorophyta were mainly present in the bare tailing, while some Proteobacteria and Cercozoa were predominantly seen in the rhizosphere. Pearson correlation and Random Forest revealed the biotic factors driving soil multifunction. Structural equation modelling further revealed that pioneer plants improved soil multifunction primarily by decreasing the microbial biodiversity and increasing the multitrophic network complexity. Overall, this highlights the importance of subterrestrial organisms in accelerating soil rehabilitation during natural succession and provides options for the ecological restoration of degraded REE mining areas.

Exploring the Relationship Among Lipid Profile Changes, Growth, and Reproduction in Folsomia candida Exposed to Teflubenzuron Over Time.

The integration of untargeted lipidomics approaches in ecotoxicology has emerged as a strategy to enhance the comprehensiveness of environmental risk assessment. Although current toxicity tests with soil microarthropods focus on species performance, that is, growth, reproduction, and survival, understanding the mechanisms of toxicity across all levels of biological organization, from molecule to community is essential for informed decision-making. Our study focused on the impacts of sublethal concentrations of the insecticide teflubenzuron on the springtail Folsomia candida. Untargeted lipidomics was applied to link changes in growth, reproduction, and the overall stress response with lipid profile changes over various exposure durations. The accumulation of teflubenzuron in organisms exposed to the highest test concentration (0.035 mg a.s. kg-1 soil dry wt) significantly impacted reproductive output without compromising growth. The results suggested a resource allocation shift from reproduction to size maintenance. This hypothesis was supported by lipid shifts on day 7, at which point reductions in triacylglycerol and diacylglycerol content corresponded with decreased offspring production on day 21. The hypermetabolism of fatty acids and N-acylethanolamines on days 2 and 7 of exposure indicated oxidative stress and inflammation in the animals in response to teflubenzuron bioaccumulation, as measured using high-performance liquid chromatography-tandem mass spectrometry. Overall, the changes in lipid profiles in comparison with phenotypic adverse outcomes highlight the potential of lipid analysis as an early-warning tool for reproductive disturbances caused by pesticides in F. candida. Environ Toxicol Chem 2024;43:1149-1160. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Trophic Transfer and Toxic Potency of Rare Earth Elements along a Terrestrial Plant-Herbivore Food Chain.

Extensive rare earth element (REE) mining activities have caused REE contamination of ambient agricultural soils, posing threats to associated food webs. Here, a simulated lettuce-snail food chain was conducted to evaluate the trophic transfer characteristics and the consequent effects of REEs on consumers. After 50-day exposure to soil, lettuce roots dose-dependently accumulated 9.4-76 mg kg-1 REEs and translocated 3.7-20 mg kg-1 REEs to shoots. Snails feeding on REE-contaminated shoots accumulated 3.0-6.7 mg kg-1 REEs with trophic transfer factors of 0.20-0.98, indicating trophic dilution in the lettuce-snail system. REE profiles in lettuce and snails indicated light REE (LREE) enrichment only in snails and the varied REE profiles along the food chain. This was corroborated by toxicokinetics. Estimated uptake (Ku) and elimination (Ke) parameters were 0.010-2.9 kgshoot kgsnail-1 day-1 and 0.010-1.8 day-1, respectively, with higher Ku values for LREE and HREE. The relatively high Ke, compared to Ku, indicating a fast REE elimination, supports the trophic dilution. Dietary exposure to REEs dose-dependently affected gut microbiota and metabolites in snails. These effects are mainly related to oxidative damage and energy expenditure, which are further substantiated by targeted analysis. Our study provides essential information about REE bioaccumulation characteristics and its associated risks to terrestrial food chains near REE mining areas.

Comments to "Degli-Innocenti, F. The pathology of hype, hyperbole and publication bias is creating an unwarranted concern towards biodegradable mulch films" [J. Hazard. Mater. 463 (2024) 132923].

Some narratives present biodegradable plastic use for soil mulching practices in agriculture as "environmentally friendly" and "sustainable" alternatives to conventional plastics. To verify these narratives, environmental research recently started focusing on their potential impact on soil health, highlighting some concerns. The paper by Degli-Innocenti criticizes this unfolding knowledge arguing that it is affected by communication hypes, alarmistic writing and a focus on exposure scenarios purposedly crafted to yield negative effects. The quest of scientists for increased impact - the paper concludes - is the driver of such behavior. As scholars devoted to the safeguarding of scientific integrity, we set to verify whether this serious claim is grounded in evidence. Through a bibliometric analysis (using number of paper reads, citations and mentions on social media to measure the impact of publications) we found that: i) the papers pointed out by Degli-Innocenti as examples of biased works do not score higher than the median of similar publications; ii) the methodology used to support the conclusion is non-scientific; and iii) the paper does not fulfil the requirements concerning disclosure of conflicts of interests. We conclude that this paper represents a non-scientific opinion, potentially biased by a conflict of interest. We ask the paper to be clearly tagged as such, after the necessary corrections on the ethic section have been made. That being said, the paper does offer some useful insights for the definition of exposure scenarios in risk assessment. We comment and elaborate on these proposed models, hoping that this can help to advance the field.

Source-specific probabilistic risk assessment of microplastics in soils applying quality criteria and data alignment methods.

The risk characterization of microplastics (MP) in soil is challenging due to the non-alignment of existing exposure and effect data. Therefore, we applied data alignment methods to assess the risks of MP in soils subject to different sources of MP pollution. Our findings reveal variations in MP characteristics among sources, emphasizing the need for source-specific alignments. To assess the reliability of the data, we applied Quality Assurance/Quality Control (QA/QC) screening tools. Risk assessment was carried out probabilistically, considering uncertainties in data alignments and effect thresholds. The Hazardous Concentrations for 5% (HC5) of the species were significantly higher compared to earlier studies and ranged between 4.0 × 107 and 2.3 × 108 particles (1-5000 µm)/kg of dry soil for different MP sources and ecologically relevant metrics. The highest risk was calculated for soils with MP entering via diffuse and unspecified local sources, i.e., "background pollution". However, the source with the highest proportion of high-risk values was sewage, followed by background pollution and mulching. Notably, locations exceeding the risk threshold obtained low scores in the QA/QC assessment. No risks were observed for soils with compost. To improve future risk assessments, we advise to primarily test environmentally relevant MP mixtures and adhere to strict quality criteria.

Biotic Ligand Modeling for the Effect of Major Cations on the Uptake of Cadmium in Folsomia candida Exposed in a Sand-Solution Medium.

Biotic ligand modeling (BLM) approaches are already applied to predict the bioavailability and possible risk of metals in surface water, but need further development for soils. The present study investigated the effect of major cations (Ca2+, Mg2+, Na+, K+, and H+) on cadmium bioaccumulation in the springtail Folsomia candida. To avoid the complexity of real soils and enable control of elemental speciation in the exposure medium, the animals were exposed to different cadmium concentrations in an inert quartz sand-solution medium. Accumulation of cadmium in the animals was measured after 7 days exposure at different cation concentrations. Among the cations, only Ca2+ significantly affected the uptake of cadmium in the springtails. Mg2+ also had higher effects compared with other selected cations. Using a BLM approach, the uptake of cadmium in the animals predicted by taking into account both Ca2+ and Mg2+ activities correlated well with the measured values (R2 = 0.68). The final estimated conditional binding constants for cadmium (log KCd-BL), Ca (log KCa-BL), and Mg (log KMg-BL) of 1.06, 2.14, and 1.23 L/mol, respectively, were in agreement with previously reported values. The match between predicted and measured uptake data confirms the applicability and usefulness of the BLM for predicting the bioavailability of cadmium to springtails and opens the way for its application in soil. Environ Toxicol Chem 2024;43:1090-1096. © 2024 SETAC.

Maternal exposure to polystyrene nanoplastics causes defective retinal development and function in progeny mice by disturbing metabolic profiles.

Microplastics (MPs) and nanoplastics (NPs) are widely spreading in our living environment, accumulating in the human body and potentially threating human health. The retina, which is a terminally differentiated extension of the central nervous system, is essential for the visual system. However, the effects and molecular mechanisms of MPs/NPs on retina development and function are still unclear. Here, we investigated the effects and modes of action of polystyrene NPs (PS-NPs) on the retina using mice as a mammalian model species. Maternal PS-NP exposure (100 nm) at an environmentally realistic concentration of 10 mg L-1 (or 2.07 *1010 particles mL-1) via drinking water from the first day of pregnancy till the end of lactation (21 days after birth) caused defective neural retinal development in the neonatal mice, by depositing in the retinal tissue and reducing the number of retinal ganglion cells and bipolar cells. Exposure to PS-NPs retarded retinal vascular development, while abnormal electroretinogram (ERG) responses and an increased level of oxidative stress were also observed in the retina of the progeny mice after maternal PS-NP exposure. Metabolomics showed significant dysregulation of amino acids that are pivotal to neuron retinal function, such as glutamate, aspartate, alanine, glycine, serine, threonine, taurine, and serotonin. Transcriptomics identified significantly dysregulated genes, which were enriched in processes of angiogenesis, visual system development and lens development. Regulatory analysis showed that Fos gene mediated pathways could be a potential key target for PS-NP exposure in retinal development and function. Our study revealed that maternal exposure to PS-NPs generated detrimental effects on retinal development and function in progeny mice, offering new insights into the visual toxicity of PS-NPs.

Multigenerational exposure of Ag materials (nano and salt) in soil - environmental hazards in Enchytraeus crypticus (Oligochaeta).

Because of its properties, silver is among the most used metals both as salt and as nanomaterials (NMs), hence reaching the environment. Multigenerational (MG) exposure testing is scarce, and especially so for NMs and soil invertebrates. In this study the MG effects of Ag NMs (Ag NM300K) and Ag salt (AgNO3) were assessed, using Enchytraeus crypticus in LUFA 2.2 soil. Survival, reproduction and internal Ag concentration in the animals were measured throughout 7 generations (5 generations (F0-F4) in spiked soil plus 2 (F5-F6) in clean soil) exposed to sublethal concentrations corresponding to the reproduction EC10 and EC50 obtained in standard toxicity tests (45 and 60 mg Ag per kg soil DW for AgNO3; 20 and 60 mg Ag per kg soil DW for Ag NM300K). MG exposure caused a dose-related decrease in reproduction for both Ag forms. Ag uptake peaked in the F1 (64 days) for AgNO3 and F2 (96 days) for Ag NM300K, after which it decreased. In agreement with toxicokinetic studies, a maximum body Ag concentration was reached (20 mg Ag per kg body DW (AgNO3) and 70 mg Ag per kg body DW (Ag NM300K)) and after which detoxification mechanisms seem to be activated with elimination of Ag accompanied by a decrease in reproduction. Transfer to clean soil allowed Ag to be (fully) eliminated from the animals. This MG study confirmed the effects determined in standard reproduction toxicity tests but further allowed to monitor the dynamics between exposure and effects of the Ag materials, and how the animals seem to cope with Ag for 7 generations by compensating between detoxification and reproductive output.

The influence of soil organic matter content and substance lipophilicity on the toxicity of pesticides to the earthworm Eisenia andrei.

To account for potential differences in bioavailability (and toxicity) due to different soil organic matter (OM) contents in natural and artificial soil (AS), in the current European environmental risk assessment (ERA) a correction factor (CF) of 2 is applied to toxicity endpoints for so called lipophilic pesticides (i.e. log Kow > 2) generated from laboratory tests with soil invertebrates. However, the appropriateness of a single CF is questioned. To improve the accuracy of ERA, this study investigated the influence of soil OM content on the toxicity to the earthworm Eisenia andrei of five active substances used in pesticides covering a wide range of lipophilicity. Laboratory toxicity tests were performed in AS containing 10 %, 5 % and 2.5 % peat, and a natural LUFA 2.2 soil (4.5 % OM), assessing effects on survival, biomass change and reproduction. Pesticide toxicity differed significantly between soils. For all pesticides, toxicity values (LC50, EC50) strongly correlated with soil OM content in AS (r2 > 0.82), with toxicity decreasing with increasing OM content. Obtained regression equations were used to calculate the toxicity at OM contents of 10.0 % and 5.0 %. Model-estimated toxicity between these soils differed by factors of 1.9-3.6, and 2.1-3.2 for LC50 and EC50 values, respectively. No clear relationships between pesticide lipophilicity and toxicity-OM relationships were observed: the toxicity of non-lipophilic and lipophilic pesticides was influenced by OM content in a similar manner. The results suggest that the CF of 2 may not be appropriate as it is based on incorrect assumptions regarding the relationships between lipophilicity, OM content and toxicity. Further research should be conducted to understand the mechanistic link between toxicity and soil OM content to better define more chemically and ecologically appropriate CFs for ERA.

Intermediate-tier options in the environmental risk assessment of plant protection products for soil invertebrates-Synthesis of a workshop.

The European environmental risk assessment (ERA) of plant protection products follows a tiered approach. The approach for soil invertebrates currently consists of two steps, starting with a Tier 1 assessment based on reproduction toxicity tests with earthworms, springtails, and predatory mites. In case an unacceptable risk is identified at Tier 1, field studies can be conducted as a higher-tier option. For soil invertebrates, intermediate tiers are not implemented. Hence, there is limited possibility to include additional information for the ERA to address specific concerns when the Tier 1 fails, as an alternative to, for example, a field study. Calibrated intermediate-tier approaches could help to address risks for soil invertebrates with less time and resources but also with sufficient certainty. A multistakeholder workshop was held on 2-4 March 2022 to discuss potential intermediate-tier options, focusing on four possible areas: (1) natural soil testing, (2) single-species tests (other than standard species), (3) assessing recovery in laboratory tests, and (4) the use of assembled soil multispecies test systems. The participants acknowledged a large potential in the intermediate-tier options but concluded that some issues need to be clarified before routine application of these approaches in the ERA is possible, that is, sensitivity, reproducibility, reliability, and standardization of potential new test systems. The definition of suitable assessment factors needed to calibrate the approaches to the protection goals was acknowledged. The aims of the workshop were to foster scientific exchange and a data-driven dialog, to discuss how the different approaches could be used in the risk assessment, and to identify research priorities for future work to address uncertainties and strengthen the tiered approach in the ERA for soil invertebrates. This article outlines the background, proposed methods, technical challenges, difficulties and opportunities in the ERA, and conclusions of the workshop. Integr Environ Assess Manag 2024;20:780-793. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

UV Light Causes Structural Changes in Microplastics Exposed in Bio-Solids.

Bio-solids (biological sludge) from wastewater treatment plants are a significant source of the emission of microplastics (MPs) into the environment. Weakening the structure of MPs before they enter the environment may accelerate their degradation and reduce the environmental exposure time. Therefore, we studied the effect of UV-A and UV-C, applied at 70 °C, on three types of MPs, polypropylene (PP), polyethylene (PE), and polyethylene terephthalate (PET), that are commonly found in sewage sludge, using three shapes (fibers, lines, granules). The MPs were exposed to UV radiation in bio-solid suspensions, and to air and water as control. The structural changes in and degradation of the MPs were investigated using Attenuated Total Reflectance-Fourier Transform Infrared Spectrometry (ATR-FTIR) and surface morphology was performed with SEM analysis. UV exposure led to the emergence of carbonyl and hydroxyl groups in all of the PP samples. In PE and PET, these groups were formed only in the bio-solid suspensions. The presence of carbonyl and hydroxyl groups increased with an increasing exposure time. Overall, UV radiation had the greatest impact on the MPs in the bio-solids suspension. Due to the surface-to-volume ratio of the tested samples, which influences the degradation rate, the fibers were more degraded than the other two plastic shapes. UV-A was slightly more effective at degrading the MPs than UV-C. These findings show that ultraviolet radiation in combination with an elevated temperature affects the structure of polymers in wastewater bio-solids, which can accelerate their degradation.

Using dietary exposure to determine sub-lethal effects from imidacloprid in two springtail (Collembola) species.

Standard toxicity tests expose springtails (Collembola) through soil, while dietary exposure tests with animals visible on a surface are less commonly applied. We refined a method for dietary chemical exposure for two widely distributed and abundant Collembola species: Folsomia quadrioculata and Hypogastrura viatica as existing methods were sub-optimal. Newly hatched Collembola were offered bark with a natural layer of Cyanobacteria that was either moistened with a solution of the neonicotinoid insecticide imidacloprid using a micropipette or soaked in the solution overnight. The first method was superior in producing a measured concentration close to the nominal (0.21 and 0.13 mg/kg dry bark, respectively), and resulting in sub-lethal effects as expected. The adult body size was reduced by 8% for both species, but egg production only in H. viatica. Contrastingly, soaked bark resulted in a measured concentration of 8 mg/kg dry bark, causing high mortality and no egg production in either species. Next, we identified the sub-lethal concentration-range by moistening the bark to expose H. viatica to 0, 0.01, 0.04, 0.13, 0.43 and 1.2 mg imidacloprid/kg dry bark. Only the highest concentration affected survival, causing a mortality of 77%. Imidacloprid reduced moulting rate and the body size at first reproduction. The age at first reproduction appeared delayed as some replicates did not reproduce within the experiment duration. The method of moistened bark for dietary exposure proved optimal to continuously study life history traits, such as growth and reproductive outcomes, which are important to understand effects on key events crucial for population viability and growth.

Earthworm Coelomocyte Internalization of MoS2 Nanosheets: Multiplexed Imaging, Molecular Profiling, and Computational Modeling.

Fully understanding the cellular uptake and intracellular localization of MoS2 nanosheets (NSMoS2) is a prerequisite for their safe applications. Here, we characterized the uptake profile of NSMoS2 by functional coelomocytes of the earthworm Eisenia fetida. Considering that vacancy engineering is widely applied to enhance the NSMoS2 performance, we assessed the potential role of such atomic vacancies in regulating cellular uptake processes. Coelomocyte internalization and lysosomal accumulation of NSMoS2 were tracked by fluorescent labeling imaging. Cellular uptake inhibitors, proteomics, and transcriptomics helped to mechanistically distinguish vacancy-mediated endocytosis pathways. Specifically, Mo ions activated transmembrane transporter and ion-binding pathways, entering the coelomocyte through assisted diffusion. Unlike molybdate, pristine NSMoS2 (P-NSMoS2) induced protein polymerization and upregulated gene expression related to actin filament binding, which phenotypically initiated actin-mediated endocytosis. Conversely, vacancy-rich NSMoS2 (V-NSMoS2) were internalized by coelomocytes through a vesicle-mediated and energy-dependent pathway. Mechanistically, atomic vacancies inhibited mitochondrial transport gene expression and likely induced membrane stress, significantly enhancing endocytosis (20.3%, p < 0.001). Molecular dynamics modeling revealed structural and conformational damage of cytoskeletal protein caused by P-NSMoS2, as well as the rapid response of transport protein to V-NSMoS2. These findings demonstrate that earthworm functional coelomocytes can accumulate NSMoS2 and directly mediate cytotoxicity and that atomic vacancies can alter the endocytic pathway and enhance cellular uptake by reprogramming protein response and gene expression patterns. This study provides an important mechanistic understanding of the ecological risks of NSMoS2.

Daphnia magna Multigeneration Exposure to Carbendazim: Gene Transcription Responses.

The world population is experiencing colossal growth and thus demand for food, leading to an increase in the use of pesticides. Persistent pesticide contamination, such as carbendazim, remains a pressing environmental concern, with potentially long-term impacts on aquatic ecosystems. In the present study, Daphnia magna was exposed to carbendazim (5 µg L-1) for 12 generations, with the aim of assessing gene transcription alterations induced by carbendazim (using a D. magna custom microarray). The results showed that carbendazim caused changes in genes involved in the response to stress, DNA replication/repair, neurotransmission, ATP production, and lipid and carbohydrate metabolism at concentrations already found in the environment. These outcomes support the results of previous studies, in which carbendazim induced genotoxic effects and reproduction impairment (increasing the number of aborted eggs with the decreasing number of neonates produced). The exposure of daphnids to carbendazim did not cause a stable change in gene transcription between generations, with more genes being differentially expressed in the F0 generation than in the F12 generation. This could show some possible daphnid acclimation after 12 generations and is aligned with previous multigenerational studies where few ecotoxicological effects at the individual and populational levels and other subcellular level effects (e.g., biochemical biomarkers) were found.

A toxicokinetics approach using Enchytraeus crypticus to evaluate the efficiency of hydroxyapatite to remediate soils contaminated with rare earth elements.

Extensive rare earth element (REE) mining activities pose threats to agricultural soils surrounding the mining areas. Here, low and high REE-contaminated soils from farmlands around mine tailings were remediated with hydroxyapatite. A toxicokinetic approach was applied to assess whether the use of hydroxyapatite reduced the bioavailability of REEs and thus inhibited their accumulation in the terrestrial organism Enchytraeus crypticus. Our results showed that addition of hydroxyapatite increased soil pH, DOC and anion contents. CaCl2-extractable REE concentrations significantly decreased, indicating the stabilization by hydroxyapatite. The influence of hydroxyapatite on the REE accumulation in enchytraeids was quantified by fitting a toxicokinetic model to dynamic REE body concentrations. The estimated uptake (Ku) and elimination rate constants (Ke), and bioaccumulation factor (BAF) for REEs were in the range of 0.000821 - 0.122 kgsoil/kgworm day-1, 0.0224 - 0.136 day-1, and 0.00135 - 1.96, respectively. Both Ku and BAF were significantly reduced by over 80% by hydroxyapatite addition, confirming the decreased REE bioavailability. Low atomic number REEs had higher BAFs in slightly contaminated soil, suggesting a higher bioaccumulation potential of light REEs in soil organisms. Overall, chemical stabilization with amendments can attenuate the bioavailability of REEs and reduce the potential ecological risk of contaminated agricultural soils near REE mining areas.